NL1025709C2 - 3- (1-3- (1,3-benzothiazol-6-yl) propylcarbamoylcycloalkyl) propanoic acid derivatives as fake inhibitors. - Google Patents

Description

3- (1- [3- (1/3-BENZOTHIAZOL-6-YL) PROPYLCARBAMOYL] CYCLOALKY1 ·) PROPANIC VIRGIN DERIVATIVES AS

FAKE BRAKES

The present invention relates to inhibitors of neutral endopeptidase enzyme (NEP), uses thereof, methods for their preparation, intermediates used in their preparation, and compositions containing such inhibitors. These inhibitors are useful in all kinds of therapeutic areas, including the treatment of male and female sexual dysfunction, in particular female sexual dysfunction (female sexual dysfunction, FSD), especially when the PSD female sexual arousal disorder arousal disor-der, FSAD).

NEP inhibitors are described in WO 91/07386, WO 91/10644, WO 02/02513, WO 02/079143 and EP 1,258,474.

The use of NEP inhibitors for treating FSD is described in EP 1,097,719-A1. The use of NEP inhibitors for treating male sexual dysfunction (MSD) is described in WO 02/03995.

The present invention provides a class of potent NEP inhibitors, which have the advantage that they are selective for NEP over soluble secreted endopeptidase (SEP). The compounds of the present invention are also selective for NEP over ACE. In addition to their selectivity, the compounds of the present invention also have unexpectedly good pharmaceutical 1025709

2 I

cokinetic properties, in particular good I

oral bioavailability and a suitable method

.king time for the useful effect in νίνο. I

According to a first aspect, the present I

Invention in a compound of the general formula (I) I

or pharmaceutically acceptable salts, solvates or polymorphs

I

in 1 I-I

p ’O Η Γ Τή · I

0 0 0) I

15 wherein: I

R 1 is H or CH 3, -I

R2 is C1 -C2 alkyl and I

η is 1 or 2. I

A preferred aspect of the present invention I

20 are compounds of formula (I) wherein η is 1. I

In another preferred embodiment, R 1 is methyl. I

In yet another preferred embodiment, R2 is me-I

thyl. I

An embodiment of the present invention that I

Compounds of formula I are particularly preferred

le (I), wherein R1 is methyl, R2 is methyl and η is 1; R1 was wa

is substance, R2 is ethyl and η is 1; R1 is methyl, R2 is ethyl I

and η is 1; and R 1 is hydrogen, R 2 is ethyl and n is 2. I

The compounds of the present invention are: I

30 (1) (R) -2-Methyl-3- (1 - {[3- (2-methyl-1,3-benzothiazole-6-)

yDpropyl] carbamoyl} cycl opens yl) propanoic acid (Example 1), I

(2) 3- (1 - {[3- (2-ethyl-1,3-benzothiazole-6-l)

yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 2), I

(3) (R) -2-Methyl-3- (1 - {[3- (2-ethyl-1,3-benzothiazole-6-)

35 yDpropyl] carbamoyl} cyclopentyl propanoic acid (Example 4), I

1025709 I

3 (4) 3- (1 - {[3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid (Example 3), (5) 3- (1- {[ 3- (2-Methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid (Example 5), 5 (6) 3- (1 - {[3- (2-Methyl-1,3) -benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 6), (7) (R) -2-Methyl-3- (1 - {[3- (2-methyl-1,3-benzothiazole) 6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid and (8) (R) -2-Methyl-3- (1 - {[3- (2-ethyl-1,3-benzothiazol-6-10 yl) propyl] carbamoyl} cyclohexyl) propanoic acid.

Preferred compounds of the present invention are: (1) (R) -2-Methyl-3- (1 - {[3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid ( Example 1), 15 (2) 3- (1 - {[3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 2), (3) (R) -2-Methyl-3- (1- {[3- (2-ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 4), (4) 3- (1- { [3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid (Example 3), (5) 3- (1 - {[3- (2-Methyl-1, 3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid (example 5) and (6) 3- (1 - {[3- (2-Methyl-1,3 benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 6).

The most preferred compounds are: (1) (R) -2-Methyl-3- (1 - {[3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 1), (2) 3- (1- {1,3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 2), (3) (R) -2-Methyl-3- (1 - {[3- (2-ethyl-13-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 4) and 1025709

I 4 I

I (4) 3- (1 - ((3- (2-Ethyl-1,3-benzothiazole-6-l

III) propyl] carbamoyl} cyclohexylpropanoic acid (Example 3). I

Pharmaceutically acceptable salts of the compounds I

I of formula (I) include the acid addition and base salts I

15 (including di-salts) thereof. I

Suitable acid addition salts are formed with acids I

I which form non-toxic salts. Examples include the I

Acetate, aspartate, benzoate, besylate, bicarbonate

I / carbonate, bisulfate, camsylate, citrate, edisylate, I

Iylate, fumarate, glucepate, gluconate, glucuronate I

I, hibenzate, hydrochloride / chloride, hydrobromide I

I / bromide, hydroiodide / iodide, hydrogen phosphate, I

I isethionate, D and L lactate, malate, maleate, malo

Iate, mesylate, methyl sulfate, 2-napsylate, nicotine

Iate, nitrate, orotate, pamoate, phosphate, saccharate I

I, stearate, succinate, sulfate, D and L tartrate and I

I tosylate salts. I

Suitable base salts are formed from bases that I

I form non-toxic salts. Examples include the al

Minium, ammonium, arginine, benzathine, calcium, chocolate

Iline, diethylamine, diolamine, glycine, lysine, mag

nesium, meglumine, olamine, potassium, sodium, trom

I thamine and zinc salts. I

I For an overview of suitable salts, see: Stahl and I

Wermuth, Handbook of Pharmaceutical Salts: Properties, Se-I

Iection and Use, Wiley-VCH, Weinheim, Germany (2002). I

A pharmaceutically acceptable salt of a compound

I thing of formula (I) can easily be prepared by I

I together mixing solutions of the compound with Form

Mula (I) and the desired acid or base, to I

Depending on what applies. The salt can be from I

I precipitate the solution and are collected by filtration, or I

I are obtained by evaporation of the solvent. I

Pharmaceutically acceptable solvates in accordance with I

In accordance with the present invention, hydrates and I

1025709 solvates in which the solvent for the crystallization can be isotopically substituted, e.g. DaO, acetone-d 6, DMSO-d 6.

Also within the scope of the present invention are clathrates, drug-acceptor-inclusion complexes in which, unlike the aforementioned solvates, the drug and the acceptor are present in non-stoichiometric amounts. For an overview of such complexes, see: J. Pharm. Sci., 64, 10 (8), 1269-1288, by Haleblian (August 1975).

All references herein to compound and formula (I) include references to salts thereof and to solvates and clathrates of compounds of formula (I) and salts thereof.

The present invention includes all polymorphs of compounds of formula (I) as hereinbefore defined.

Also within the scope of the present invention are the so-called "prodrugs" of the compounds of formula (I). Certain derivatives of compounds of formula (I) which themselves have little or no pharmacological activity can, when they are metabolized after administration in or to the body, thus lead to compounds of formula (I) having the desired activity. Such derivatives are called "prodrugs".

Prodrugs in accordance with the present invention can be produced, for example, by replacing suitable functions present in the compounds of formula (I) with certain residues known to those skilled in the art as "pro-residues" and for example described in " Design of Prodrugs "by H. Bund-gaard (Elsevier, 1985).

Finally, certain compounds of formula (I) may themselves act as prodrugs of other compounds of formula (I).

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6 I

Compounds of formula (I) that have one or more asymmetric I

carbon atoms can be as two or more I

optical isomers exist. When a connection with I

formula (I) contains an alkenyl or alkenylene group, I

Geometric cis / trans (or Z / E) isomers possible, and I

where the compound is, for example, a keto or oxime group

tautomeric isomerism ("tautomerism") may occur. The I

It will be appreciated that a single connection has more than one I

type of isomerism. I

Included in the scope of the present invention I

are all optical isomers, geometric isomers, and tau-I

tomere forms of compounds of formula (I), including I

compounds that exhibit more than one type of isomerism, and I

mixtures of one or more thereof. I

Cis / trans-i sumers can be separated by I

using conventional techniques for experts I

are known, for example fractional crystallization I

and chromatography. I

Conventional preparation / isolation techniques I

20 of individual stereoisomers include the conversion I

from a suitable optically pure precursor, separation of I

the racemate (or the racemate of a salt or derivative) I

using, for example, chiral HPLC, or fractional

crystallized crystallization of diastereoisomeric salts which I

25 are formed by reacting the racemate with a gel

suitable optically active acid or suitable optic action I

ve base, for example tartaric acid. I

The present invention also includes all pharmaceuticals

Iotically acceptable isotopic variations of a compound

Thing of formula (I). An isotopic variation is observed

defined as one in which at least one atom has been replaced

gene through an atom with the same atomic number, but with an I

atomic mass that differs from the atomic mass that is usually I

is found in nature. I

1025709 I

7

Examples of isotopes suitable for incorporation into the compounds of the present invention include isotopes of hydrogen such as aH and 3H, carbon such as 13 C and 14 C, nitrogen such as 1 SN, oxygen such as 170 and 180, phosphorus such as 32 P, sulfur such as 35 S, fluorine such as ieF and chlorine such as 36Cl.

Substitution of the compounds of the present invention with isotopes such as deuterium, i.e., 2H, may provide certain therapeutic benefits that result from greater metabolic stability, for example, increased in vivo half-life or lower dosage requirements, and thus may occur in circumstances are preferred.

Certain isotopic variations of the compounds of formula (I), for example those in which a radioactive isotope is incorporated, are useful in drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e., 3 H, and carbon-14, i.e., 14 C, are particularly useful for this purpose in view of the ease of their incorporation and the easy method of detection.

Isotopic variations of the compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by methods analogous to those described in the accompanying examples and preparations using the correct isotopic variations of suitable reagents.

The compounds of formula (I) can be freeze-dried, spray-dried or dried under evaporation to provide a solid layer, a powder or a film of crystalline or amorphous material. For this purpose dr. may be used by means of microwave or radio frequency waves. .

1025709

I 8 I

The compounds of formula (I) can be obtained by means of I

I of the following process are prepared as described

In the diagram below (I): I

I e1 ƒ- (PH2) n "I

I! "V-H-"

I oo 0 0 du) / I

I w. I

I 10 I

L (f2, n I

I! U h in V-rz I

I 0 O (IV) I

15 (b). I

I R1 Γ (ξΗ *) η I

I § Lx h \ r ^ —r2 i

I 0 0 (η I

I 20 w I

The compounds of formula (IV) can be prepared I

By reacting compounds of formula (II)

I and (III) under the conditions of step (a). of the method: I

Formation of amide bond - such reactions can

I 25 of all kinds of conditions are carried out that for experts

Are known. I

In a typical case, the carboxylic acid can be activated

I is initiated by treatment with an agent such as 1,1'-I

Carbonyldiimidazole (GDI), fluoro-Ν, Ν, Ν ', Ν'-tetramethyl-I

Formamidinium hexafluorophosphate (TFFH), or a combination I

I of reagents such as azabenzotriazole-1-yloxytris (pyrr

I lidino) phosphonium hexafluorophosphate (PyAOP) and 1-hydroxy-7- I

Azabenzotriazole (HOAt). Alternatively, the reaction I

I are carried out by adding a coupling agent I

For peptides such as O- (7-azabenzotriazol-1-yl) -Ν, Ν, Ν ', N'-I

, 1025709 I

9 uronium hexafluorophosphate (HATU) or O-benzotriazol-1-yl-N, N, N ', N' uronium hexafluorophosphate (HBTU) or N, N'-dicyclohexylcarbodiimide (DCC), 1,3-diisopropylcarbodiimide (DIC) on a mixture of the acid 5 and the atnine. The reaction is carried out and a suitable solvent such as CH 2 Cl 2, pyridine, N, N-dimethylformamide (DMF), Ν, Ν-dimethylacetamide (DMA) or 1-methyl-2-pyrrolidinone between 0 ° C and the boiling point of the solvent.

The reaction is preferably carried out with the aid of CDI, triethylamine and isopropyl acetate as a solvent.

The product of step (a) of the process is then treated under the conditions of step (b) of the process: removal of the protecting group BG. Suitable groups are described in "Protective Groups in Organic Synthesis" by T.W. Greene and P.G.M. Wuts, John Wiley and Sons, Ine., 1991.

The conditions required for removal of the protecting group are often specific to that protecting group; conditions for their removal can be found in references such as Greene, T.W., Wuts, P.G.M., Protective Groups in Organic Synthesis, Wiley Interscience, and Kocienski, P.J., Protecting Groups, Thieme.

O

Preferably, BG is a tert-butyl group and the deprotection consists of acid catalysis with a suitable solvent at room temperature. The preferred conditions are trifluoroacetic acid in dichloromethane.

R, r-ipH *) n 30 / PG-0Y> X> OH + +. (II) (b) .35 1 0 25 7 0

10 I

Ρ'Γ (\ Η2) "I

? L) η | Γ V-r2 I I

χ · ^^ - ε! I

O o (VI) I

(X) I (a) I

1 r ^? Hz) n I

? L) h fTv * 2 I

PQ-OyN ^ YNv ^ V ^ s I

o o (IV) I

In an alternative method, compounds with I

formula (IV) are prepared from compounds of formula

(X) and (VI) by step (a) of the process comprising a coupling

Aryl-alyl. Suitable conditions are for I

the experts known. The compounds of formula (X)

can be prepared from compounds of formula (II) and I

allylamine by step (b) of the process which forms

an amide bond. Such reactions can be described under I

20 all kinds of conditions are carried out that for experts I

be famous.

The compounds of formula (II) and (III) can be I

prepared according to methods described in WO I

02/079143. I

In addition, compounds of formula (III) can be used

prepared as follows: I

e

(V) I

(VI)

30 I

(C) I

35 I

1025709 I

11 PG, ^ 'νΙ'Ι PG s (VII) (t)

5 t ^ r ^ K

λ ιΐΓ y — r2 (fli)

The compounds of formula (VII) can be prepared from compounds of formula (V) and (VI) under the conditions of step (c) of the process, a coupling of aryl-allyl. Suitable conditions are known to those skilled in the art. Particularly suitable conditions are those of the coupling reaction of Suzuki-Miyaura [Angew. Chem. Int. Ed., 2001, 40 (24), 4544-4568], with a hydrogenated intermediate prepared from a suitably protected allylamine derivative (e.g. di (tert-butyl) allyl imidodicarbonate; Bio-organic & Medicina Chemistry Letters, 1999, 7, 1625-1636), and a borane derivative, such as 9-BBN.

The compounds of formula (III) can be prepared from compounds of formula (VII) under the conditions of step (b) of the process: removal of the protecting group BG, as described herein.

+ j ^ YA— *. (VI) (b) 30 w 35.

1025709

12 I

(VIII) I

(a) I

% -R2 I

(lil) I

Alternatively, amine (III) can be prepared from I

compounds of formula (VIII) through step (a) of the process

way that reduction of carbon-carbon multiple and I

carbon-nitrogen bonds. Suitable conditions I

are known to those skilled in the art. In particular suitable con

Ditions include treatment with Boc20, NiCl2 and NaBK1, gel

followed by deprotection of the resulting tertiary butyl I

group as described above. The compounds with I

formula (VIII) can be prepared from acrylonitrile and I

compounds of formula (VI) through step (b) of the working I

Manner comprising coupling of aryl-allyl, wherein X is a ha

is, preferably iodine. Suitable conditions are I

known to the experts. In particular suitable conditions I

include treatment with Pd (OAc) 2 <P (o-tolyl) 3 and NaOAc in I

DMF. / v. Kl I

5 I

25 yr. I / _ "R2 I

o I

(IX) I

30 I (a) I

35

(VIII) I

1 025 7G 9 I

13

Alternatively, the compounds of formula (VIII) may be prepared from cyanoacetic acid and compounds of formula (IX) by step (a) of the process: condensation. Suitable conditions are known to those skilled in the art. Suitable conditions are described in "Advanced Organic Chemistry" by Jerry March, 4th edition, Wiley-Interscience, page 945. The compounds of formula (IX) are known in the literature (e.g., Zhurnal Obshchei Khi-mii, (1964) , 34 (11), 3801-6).

All of the above reactions and the preparations of new starting materials used in the foregoing processes are conventional. Suitable reagents and reaction conditions for their performance or preparation, as well as procedures for isolating the desired compounds, are known to those skilled in the art, with reference to literature precursors and the examples and preparations below.

The compounds of the present invention form a class of NEP inhibitors that are selective for NEP over SEP.

In general, it is important for a drug to be as selective as possible for the desired target enzyme; additional activities give rise to possible side effects. SEP has been identified relatively recently and its exact physiological role still needs to be determined in full. Regardless of what role SEP may or may not play, it is, however, a medical rule to ensure that a drug has selectivity over closely related mechanistic goals of an unknown physiological function.

Without being bound by any theory, it becomes clear that NEP and SEP are capable of hydrolysis 1025709

14 I

of many of the same biologically important peptides I

such as enkephalin, endothelin (ET), big endothelin (Big I

ET), bradykinin, substance P, angiotensin 1, atrial I

natriuretic peptide (ANP) and gonadotropic releasing hor-I

5 mone, (GnRH). - I

If a patient is being treated with a medicine I

that inhibits NEP and SEP, the hydrolysis of many of them

they are peptides (most of which are not involved in the I

improvement of the sexual function associated with I

10 the inhibition of NEP) will be reduced and the levels I

of these peptides therefore increase. A number of updating

in connection with an increase in the levels of this I

peptides can be assumed: the blood pressure can be lower I

when the ANP levels rise; an increase in I

The single-phalin levels can lead to changes in the I

pain perception; endothelin-1 is a powerful vasocon-I

strictor that changes the degree of conversion of Big ET into ET

or the hydrolysis of ET-1 can lead to changes I

of blood pressure. I

Thus, if the patient is an inhibitor of NEP I

given, the increases in the levels of this I

substrate peptides are less since the active SEP-I

enzyme will still be present. All side effects in I

related to changes in the levels of these peptides I

25 will therefore also be less. I

In addition, the mRNA can be used for SEP in various genes

stops are found in other tissues, including I

the testes, the heart, the brain, the kidneys, the saliva I

glands, thyroid glands, placenta, small intestine and I

30 the ovary (own data and from Bonvouloir et al.). I

In mice, the RNA for SEP has also been demonstrated in the spleen I

and the adrenal glands. A NEP inhibitor that does not inhibit SEP will be I

therefore probably a greater chance of a cleaner I

provide a physiological profile. I

1025709 _ ___I

15

Quite surprisingly, by identifying a class of NEP retnmers that are selective with respect to SEP, it has been discovered that these compounds have favorable pharmacokinetic properties for oral administration.

5 An orally administered drug should have a good bioavailability - that is, the ability to easily pass the gastrointestinal (MD) tract - and such a metabolic rate that it is not subjected to extensive metabolism 10 MD channel enters systemic circulation. Molecules that are rapidly metabolized will have a lower bioavailability since more compound will be removed by the metabolism as it enters the systemic circulation. As soon as a drug is in the systemic circulation, the metabolic rate of conversion is also important for determining the residence time of the drug in the body - a rapid metabolism of a drug will lead to a short duration of its action.

It is thus clear that it is important for drug molecules to possess properties that make it possible to easily pass through the MD channel and still be metabolized slowly in the body.

The CACO-2 assay is a generally accepted model for predicting the ability of a particular molecule to pass through the MD channel. The molecules of the present invention run well through the CACO-2 system.

The major part of the metabolism of drug molecules generally takes place in the liver. Therefore, the use of human liver microsomes (human liver microsomes, HLM) is a generally accepted method for measuring the sensitivity of a given molecule. molecule for liver metabolism. The compounds of the present invention are stable to HLM.

1025709

16 I

Of the compounds that provide a good passage from the I

Knowing the CACO-2 system and being stable to HLM is a priority

that this has good oral bioavailability I

(good absorption in the MD channel and minimal extraction I

5 of the compound as it passes through the liver) and a long I

duration of stay in the body - sufficient to

medicinal product to be effective. I

In addition, the compounds of the present I

invention in the free acid form crystalline, so that I

10 does not have to use any salt formation and so it is particularly important

easy to handle. I

The compounds of the present invention are I

inhibitors of the zinc-dependent, neutral endopeptide

EC.3.4.24.11., and it is suspected that the reported I

The compounds of the present invention are listed below

be able to treat the disease states. This enzyme I

is involved in the degradation of several biological I

active oligopeptides, with peptide bonds on the I

amino side of hydrophobic amino acid residues are split. I

The peptides that are metabolized include atrial I

natriuretic peptide (ANP), bombesin, bradykinin, to I

the calcitonin gene related peptide, endothelins, and I

kefalins, neurotensin, substance P and vasoactive pep I

from the intestines. Some of these peptides have I

25 powerful vasodilator and neurohormone functions, diure I

energetic and sodium-based activity or intervene in I

drag effects. I

The compounds of the present invention can be I

by inhibiting neutral endopeptidase EC.3.4.24.11. I

30 thus the biological effects of biologically active peptides

the potential. The compounds have particular use

ability to treat a number of disorders, I

including hypertension, pulmonary hypertension, peripheral I

vascular disease, heart failure, angina, renal failure

35 ficiency, acute renal insufficiency, cyclical edema, I

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17 Meniere's disease, hyperaldosteronism (primary and secondary) and hypercalciuria. The term hypertension includes all diseases characterized by supranormal blood pressure, such as essential hypertension, pulmonary hyperension, secondary hypertension, isolated systolic hypertension, hypertension associated with diabetes, hypertension associated with atherosclerosis, and renovascular hypertension, and further extends to conditions in which elevated blood pressure is a known risk factor. Accordingly, the term "treatment of hypertension" includes the treatment or prevention of complications arising from hypertension, and other related comorbidities, including congestive heart failure, angina, stroke, glaucoma, renal impairment, including renal insufficiency, obesity, and metabolic diseases (including metabolic syndrome). Metabolic diseases include in particular diabetes and impaired glucose tolerance, including its complications, such as diabetic retinopathy and diabetic neuropathy.

In addition to their ability to potentiate the effects of ANF, the compounds are useful in the treatment of glaucoma. As another outcome of their ability to inhibit neutral endopeptidase EC.3.4.24.11. the compounds of the present invention may have activity on other therapeutic agents. including, for example, the treatment of menstrual disorders, premature labor pains, preeclampsia, endometryrosis and reproductive disorders (in particular male and female infertility, polycystic ovarian syndrome, failed implantation). The compounds of the present invention would also include asthma, inflammation, leukemia, pain, pain, depression, drug use, cirrhosis, epilepsy, affective disorders, dementia and geriatric confusion, obesity, and gastrointestinal 1025709

18 I

disorders (in particular diarrhea and irritable bowel I)

syndrome), wound healing (in particular diabetic and I

venous ulcers and bedsores), septic shock, the mo- I

reduction of gastric acid secretion, the treatment of I

Hyperreninism, cystic fibrosis, restenosis, diabetic I

to treat complications and atherosclerosis. I

In a preferred embodiment, the compounds are I

of the present invention useful in the treatment I

of male and female sexual dysfunction. The

Compounds of the present invention are in particular I

beneficial for the treatment of PSD (in particular I

FSAD) and male sexual dysfunction (in particular I

male erectile dysfunction (MED). I

In accordance with the present invention, I

FSD are defined as problems with or inability I

of a woman to find satisfaction in sexual expression I

Pine tree. FSD is a common expression for ver I

various divergent female sexual disorders I

(Leiblum, S. R. (1998). Definition and classification of I

20 female sexual disorders, Int. J. Impotence Res. 10, S104-I

S106; Berman, J. R., Bernvan, L. & Goldstein, I. (1999). I

Female sexual dysfunction: Incidence, pathophysiology, I

evaluations and treatment options. Urology, 54, 385-391). I

The woman may suffer from the lack of sexual desire

25 gene, problems with excitement or orgasm, pain in I

slaughter community, or a combination of these problems. I

Various types of the disease, medications> injuries or I

psychological problems can cause FSD. Treatment I

The changes in their development form the objective to

30 specific subtypes of FSD, mainly desire and excitement disorders.

The FSD categories can best be defined

by these as opposed to the phases of a norm

male sexual response: desire, op- I

35 winding and orgasm (Leiblum, S.R. (1998). Definition and I

1025709 I

19 classification of female sexual disorders. Int. J. Imposition Res. 10, S104-S106). Desire or libido is the. incitement to sexual expression. The expressions often include sexual thoughts when one is in the company of an interested partner, or when one is exposed to other erotic stimuli. Excitement is the vascular response to sexual stimulation, an important part of which is a genital response, and this includes increased vagina moisture, vagina extension, and increased genital sensation / sensitivity.

Orgasm is the release of sexual tension built up during the excitement.

Therefore, FSD occurs when a woman has an inadequate or unsatisfactory response in one of these phases, usually desire, excitement, or orgasm. FSD categories include hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorders, and sexual pain disorders. Although the compounds of the present invention will improve the genital response to sexual stimulation (such as in female sexual arousal disorder), they may thereby also eliminate pain, problems and discomfort associated with sexual intercourse, and thus other female treat sexual disorders; 25 Hypoactive sexual desire disorder occurs when a woman has little or no desire for sex, or has no or few sexual thoughts or fantasies.

This type of FSD can be caused by low testosterone levels, due to the natural menopause or menopause caused by surgery. Other causes include illness, medication, fatigue, depression, and anxiety. .

Female sexual arousal disorder (FSAD) is characterized by inadequate genital response to sexual stimulation. The genitals do not undergo the response that 1025709

I 20 I

I features a normal sexual arousal. The walls of the I

I vagina hardly get wet, so that genitalia

I board is painful. Orgasms could be involved

I seems, The excitement disorder can be caused by I

A reduced estrogen level at the menopause or after the I

I birth of children or during nursing, as well as by I

I diseases, with vascular components such as diabetes and I

I atherosclerosis. Other causes arise from the I

I treatment with diuretics, antihistamines, antidepressants I

I 10 (e.g., SSRIs) or antihypertensive agents. I

I Sexual pain disorders (e.g. dyspareunia and vagi I

Iism) are characterized by pain that results I

I of penetration and can be caused by medication I

I moisturizing, endometriosis, inflammatory

I 15 disease of the pelvis, inflammatory disease of the dar- I

I men and urinary tract problems. I

I The extent of the occurrence of FSD is difficult to estimate I

I because the expression has several types of problems

I strokes, some of which are difficult to measure, and because I

I 20 the interest in treating FSD is a relative one

I is recent. The sexual problems of many women depend on I

I often directly together with the female aging I

I process or with chronic diseases such as diabetes and hyper-I

I tension. I

Because FSD consists of several subtypes that symbol I

I in separate phases of the sexual response cycle

I show, there is no single therapy. The current I

I treatment of FSD focuses primarily on psychology

Ical or relational matters. The treatment of FSD developed I

I gradually settles while more clinical and scientific

Basic scientific studies focusing on research by I

I address this medical problem. Female sexual I

I complaints do not all have a psychological pathophysiology

I gie, in particular for those persons who are part of the I

I 35 general female sexual complaint vasculogeneic dysfunc I

I 1025709

21 (e.g. FSAD). There are currently no authorized medicines for the treatment of FSD. Drug empirical therapy includes the administration of estrogens (topically or as a hormone replacement therapy), androgens or mood-altering drugs such as buspiron or trazodone. These treatment options are often unsatisfactory due to their limited effect or unacceptable side effects.

Since the interest in pharmacologically treating FSD is relatively recent, the therapy consists of the following: psychological advice, sexually available sex lubricants, and candidates for research, including drugs that are authorized for other conditions. These medications consist of hormonal agents, either testosterone or combinations of estrogen and testosterone. More recently, vascular drugs have been shown to be effective in male erectile dysfunction. However, none of these agents has been shown to be particularly effective in treating FSD.

The Diagnostic and Statistical Manual (DSM) IV of the American Psychiatry Association defines female sexual arousal disorder (FSAD) as: "a persistent or recurring inability to attain or maintain sexual adequacy until the end of sexual activity swelling with sexual arousal. The disorder causes considerable discomfort or problems between people. "

The excitement response consists of vasocongestion in the pelvis, moistening of the vagina and expansion and swelling of the external genitalia. The disorder causes considerable discomfort and / or problems between people.

FSAD is a very common sexual disorder that affects pre-, peri- and postmenopausal (+ HRT) women.

1025709

I 22 I

I This is accompanied by concomitant disorders, such as I

I depression, cardiovascular diseases, diabetes and UG-I

I disorders. I

I The main consequences of FSAD are the lack I

I 5 from a response / swelling, not getting moist enough and I

I the lack of a pleasant genital sensation. The se

I secondary effects of FSAD are reduced sexual desire, I

I pain during sexual intercourse and problems with I

I achieve an orgasm. I

I 10 It has recently been assumed that there is a vascular ground I

I exists for at least a part of the patients with I

I symptoms of FSAD (Goldstein et al., Int. J. Impot. Res., I

I 10, S84-S90, 1998), with data from animals this I

I support vision (Park et al., Int. J. Impot. Res., 9, I

I 15 27-37, 1997). I

Medicines that are candidates for treatment

I from FSAD, the efficiency of which is being investigated, are I

I therapies for primary erectile dysfunction affecting the circle

I promote circulation in the male genitalia. This i

20 consist of two types of preparations, oral or sublingual

Medicines (Apomorphine, Phentolamine, inhibitors of phospho-I

I diesterase-5 (PDE-5), e.g. Sildenafil) and prostaglandin I

I (PGEj) that are injected or trans-tally added

I served to men, and topically to the genitals of I

I 25 women. I

The compounds of the present invention are I

I advantageous in that it provides means for the repeat I

I suggest a normal sexual arousal response - namely I

I increased blood flow of the genitals, which I

I 30 leads to a vaginal, clitoral and labial response. This I

I will result in an increased wetness of the vessel

I gina through transudation of plasma, an increased adaptation I

I of the vagina and increased genital sensitivity. The I

Thus, compounds of the present invention provide I

1025709 23 means for restoring or potentiating the normal sexual arousal response.

Without considering ourselves bound by theory, we assume that neuropeptides such as vasoactive intestinal peptide 5 (vasoactive intestinal peptide, VIP) are the main neuro-transmitter candidates for the control of the female sexual arousal response, in particular for the control of genital blood flow. VIP and other neuropeptides are degraded / metabolized by NEP EC 10 3.4.24.11. Thus, NEP inhibitors will potentiate the endogenous relaxant effect on the vasculature by VIP released during the excitation. This will lead to a treatment for FSAD, such as an increased genital blood flow and therefore a genital response. We have shown that selective inhibitors of NEP EC 3.4.24.11. reinforce pelvic nerve-induced and VIP-induced increases in vaginal and clitoral blood flow. In addition, selective NEP inhibitors enhance VIP and nervous stimulation of the insulated vaginal wall.

The present invention is thus advantageous in that it provides means for restoring a normal sexual arousal response - namely, increased blood flow of the genitals, leading to a vaginal, clitoral, and labial response. This will result in an increased moistening of the vagina via transduction of plasma, an increased adjustment of the vagina and an increased sensitivity of the vagina. The present invention therefore provides means for restoring or potentiating the normal sexual arousal response.

Male sexual dysfunction includes male erectile dysfunction, ejaculation disorders such as premature ejaculation (PE), anorgasmia (inability to reach 1025709 t ·.

24 I

orgasm) and pleasure disorders, such as hypoactive I

sexual appetite disorder (no interest in sex). I

It will be understood that all references herein I

to treatment the curative, palliative and prophylactic I

Treatment. I

The compounds of the present invention find I

application in the following subpopulations of patients with I

FSD: young, older, pre-menopausal, peri-menopausal and I

post-menopausal women with or without hormone replacement I

10 therapy. I

The compounds of the present invention find I

application in patients with FSD that results from: I

i) vasculogeneic etiologies, e.g. cardiovascular or I

atherosclerotic diseases, hypercholesterolemia, smoking I

15 of cigarettes, diabetes, hypertension, radiation and pe

linear trauma, trauma of the ilio hypogastric pudenda I

vascular system. I

ii) neurogenic aetiologies, such as spinal injury

marrow or diseases of the central nervous system, including I

Multiple sclerosis, diabetes, Parkinsonism, cerebrovascular disease

lary accidents, peripheral neuropathies, trauma or radi

pelvic surgery. I

iii) hormonal / endocrine etiologies, such as dysfunction I

of the hypothalamic / pituitary / gonadal tract, or dys-I

Ovarian function, pancreas dysfunction, surgeon

chemical or medical castration, androgen deficiency, high circulatory I

learning levels of prolactin, e.g. hyperprolactinemia, I

natural menopause, early ovarian insufficiency, I

hyper and hypothyroidism. I

Iv) psychogenic aetiologies, such as depression, obsession

sive / compulsive disorder, anxiety disorder, postnatal disorder

pressure / "baby blues", emotional and relational problems, I

fear of failure, marital problems, dysfunctional attitudes, I

sexual phobias, religious taboo or traumatic experience I

35 gen from the past. I

1025709_ V) drug-induced sexual dysfunction that is the result of selective inhibitor therapy; re-uptake of serotonin (SSRIs) and other antidepressant therapies (with tricyclic compounds and the major tranquilizers), anti-hypertensive therapies, sympatholytic drugs, chronic therapy of oral contraceptives.

Patients with mild to moderate MED would benefit from treatment with a compound of the present invention, and patients with severe MED could also respond. However, preliminary studies suggest that the response rate of patients with mild, moderate and severe MED will be greater in combination with an inhibitor of PDE5. Mild, moderate and severe MED are expressions known to those skilled in the art, but guidelines can be found in The Journal of Urology, vol. 151, 54-61 (January 1994).

The compounds of the present invention find use in the following subpopulations of patients with MED: psychogenic, endocrine, nerogenic, atherogenic, and drug-induced sexual dysfunction (lac-togene), and sexual dysfunction associated with swelling body factors concerns, in particular venogenic causes. These patient groups are described in more detail in Clinical Andrology, vol. 23, No. 4, pp. 773-782, and Chapter 3 of the book by I. Eardley and K. Sethia "Erectile Dysfunction - Current Investigation and Management, published by Mosby-Wolfe.

30 Assay conditions

Production of natural NEP enzyme: NEP is isolated from kidneys by following the method described by Kenny and Booth (Booth, A. G. & Kenny, A. J. (1974), Biochem. J., 142, 575-581).

1025709

I 26 I

I Recombinant SEP enzyme is produced using I

I of one of the two alternative methods: I

Method 1: a culture of Chinese hamster ovary I

I (CHO) cells are transfected with the plasmid NCIMB, I

Registration number 41110, wherein the lipofectamine method I

I is used as described in the protocol for I

I lipofectamine reagent (Invitrogen Ltd., Paisley, UK). The I

I cell medium is harvested 24 or 48 hours after transfection and I

I stripped of cell debris by centrifugation for 5 minutes

Run at 3000 x g. The medium is then added during I

I dialyzed overnight at 4 ° C against 50 mM HEPES with I

PH = 7.4 / 10% glycerol, with the aid of a slider al lyser

I from Pierce and Warner, Chester, UK. The dialyzed sample

The star is then frozen in small volumes and stored

Worth 15 under liquid nitrogen. I

Method 2: a stable human embryonic kidney I

I (human embryonic kidney, HEK) cell line which is recombinant SEP I

I produces, was made in-house according to I

I molecular and cell biological standard methods. This I

HEK-SEP cell line is grown in flasks or roller bottles

I according to standard protocols for HEK cells, in media I

I supplemented with hygromycin B. The medium becomes I

I collected and stored at room temperature for 15 minutes

I centrifuged at 3000 x g to remove the cell debris I

I and then dialysed with I for at least 6 hours

I dialysis buffer (50 fflM HEPES with pH = 7.4 / 10% glycerol), where I

I with a "slide a lyser" by Pierce and Warner, Chester, UK, I

I is used. The dialysis buffer is used during these 6 hours

Replace at least once. I

I 30 I

Assay for SEP or NEP peptidase activity I

The peptidase activity of SEP or NEP is measured

I by studying the ability to proteolize I

I of the synthetic peptide substrate Rhodamine Green-Gly-I

35 Gly-dPhe-Leu-Arg-Arg-Val-Cys (QSY7) -3Ala-NH2: I

I 1025709 I

27

The reagents for the assay are first prepared as follows:

A substrate solution is prepared by diluting a 2 mM stock solution in 100% DMSO of Rhodamine-5 green-Gly-Gly-dPhe-Leu-Arg-Arg-Val-Cys (QSY7) -pAla-> NH 2 in 50 mM HEPES buffer with pH = 7.4 (Sigma, UK) to a concentration of 2 μΜ.

A small volume of the SEP or NEP enzyme described above is thawed and then diluted in 50 mM 10 HEPES, with a pH of 7.4 containing 1 EDTA-free cocktail tablet with protease inhibitor (Roche Diagnostics, UK) per 25 ml. The dilution is made with a predetermined factor that is specific to each batch of enzyme, so that 15 μΐ contains enough enzyme to convert approximately 30% of the substrate into product during the assay.

A 4% solution in DMSO is prepared which consists of 4 ml of DMSO plus 96 ml of 50 mM HEPES with pH = 7.4. A solution of product is prepared by adding 500 μΐ substrate solution to 250 μΐ enzyme solution plus 250 μΐ 20 4% solution in DMSO, and incubating at 37 ° C for 16 hours.

The assays are deployed as follows:

15 μ mic enzyme solution is added to a black microtiter plate with 384 wells, to 15 μΐ 4% solution in 25 DMSO. A similar, non-specific blank test is used as the background value, whereby the 15 μΐ of the 4% solution in DMSO also contains 40 μΜ phosphoramidone. 30 μΐ substrate solution is then added to both the assay and the blank test, and then the plate is incubated for 1 hour at 37 ° C. A fluorescence measurement is made after the incubation (Ex485 / Em538). With a BMG galaxy fluorescent reader (BMG Lab Technologies, Offenberg, Germany).

1025709

28 I

The proteolytic activity of the enzyme is similar

one with the fluorescence of the sample minus the fluorescent

the non-specific blank test performed by the back I

land value. I

5 A fluorescence measurement of 60 l can be made

μΐ product in a well of an identical microtiter plate. I

If desired, this value, together with the fluorescent I

units of the SEP assay used for the calculation I

of the percentage of the substrate that during an in- l

The 1 hour cubing period was proteolysed or before I

converting the measured rise in fluorescence to I

other useful units, such as number of ng proteolysed I

substrate / min / ml enzyme. I

With the help of the assay, the IC 50 values of I

NEP and SEP inhibitors determined: I

To determine the IC50 of SEP or NEP inhibitors (e.g. I

example phosphoramidone), several assays are performed

as described above, wherein a series of I

test concentrations of the inhibitor in the 15 μΐ solution in I

20 DMSO is included (made by the correct dilution I

of a 10 mM stock solution of inhibitor in 100% DMSO I

with 4% DMSO / 50 mM HEPES with pH = 7.4). With the help of an I

suitable standard computer program for editing I

graphical data becomes an S-shaped dose-response curve I

25 processed to a graph of log concentration of inhibitor I

against% inhibition or% activity. The IC 50 value I

is calculated as the concentration of the inhibitor that is 50% I

of the maximum inhibition. In a typical case I

For a given IC50 determination, a dosing range of I becomes

30 uses at least 10 concentrations of inhibitor that in I

sizes differ by half. I

For inhibitors that have an IC 50 outcome of less than one

give about 2 nM, the assay is repeated under change

the assay conditions in which the amount of enzyme I used

35 is reduced to approximately 1/10 & 1/20 part; the sub-I

1025709 I

29 street concentration is increased to 5 μΜ and the incubation period is extended to 3 hours. This lowers the potential limit (strong bond limit) of the assay to a level where the estimate of the IC 50 value of the compounds whose K value is in the range of 0.2-2 nM is not limited by the concentration of the enzyme.

The compounds of the present invention have been tested in the above assays. All compounds are 10 potent NEP inhibitors with an IC 50 value of <20 nanomolar and a selectivity for NEP relative to SEP of at least 1000 times.

(R) -2-Methyl-3- (1 - {[3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 1) has an activity against NEP that can be expressed as an IC50 of 1 nM and a selectivity for NEP compared to SEP of 1900 times.

The usefulness of the compounds of the present invention for treating FSD and MED can be further determined using the techniques described in WO 02/079143.

The beneficial pharmacokinetics of the compounds of the present invention can be demonstrated by the CACO-2 test. The CACO-2 assay is a generally accepted model for predicting the ability of a particular molecule to pass through the MD channel. The compounds of the present invention pass the CACO-2 well. This can be defined as follows. Compounds with an apparent permeability (Papp) -30 value in the CACO of> 5x10 -6 cm / s (at pH = 7.4) and> 15x10 -6 cm / s (at pH = 6.5) are considered a good permeability and it can be predicted that they will be well absorbed in the MD channel.

The test is carried out as described below:

1025709

I 30 I

I Cell culture I

I CACO-2 cells were inoculated with 4.0 x 10 4 cells / well

I in Falcon Multiwell® plates with 24 wells each. The cells I

I were grown in culture media consisting of at least I

I essential medium (Gibco 21090-022) that was supplemented with I

I 20% fetal bovine serum, 1% non-essential amino acids, 2 I

1 mM L-glutamine and 2 mM sodium pyruvate. The culture medium I

I was replaced three times each week and the cells were examined

I maintained at 37 ° C, 5% CO 3 and a relative humidity of I

I 90%. The permeability studies were conducted when the I

I monolayers were between 15 and 18 days old. The cells were I

Iden used between cycles 23 and 40. I

Permeability studies: I

Each test compound was prepared as a 10 mM solution

In DMSO, and 62.5 μΐ of this solution was continued

Added to 25 ml of transport buffer. Nadolol (25 I

I μΜ) was added to each well as a marker of the I

I completeness of the membranes. These solutions were I

I together with transport buffer then heated to 37 ° C. I

The transport buffer was HBSS (Hank's balanced salt solution)

Ion) with pH = 7.4 or pH = 6.5. Before the start of each study

I was washed three times with HBSS each monolayer. Trans-I

Port buffer with no added compound was received in each

I catching well, 250 μΐ on the apical side and 1 I

1 ml in the basolateral well. The study was started I

By adding drug solution to each do-I

I napkin, 250 μΐ at the apical wells and 1 ml at the I

Basolateral well. After a two-hour incubation at I

At 37 ° C, the samples were removed from all wells for I

I LC-MS-MS analysis. I

The compounds of the present invention have I

I a CACO-2 A-B value> 5. I

I 1025709 I

31

Human liver microsomes are a generally accepted model for predicting the metabolic stability of drug molecules for liver metabolism. The compounds of the present invention are stable to metabolism by HLM. Compounds with a half-life in HLM of <90 minutes are metabolised too quickly and may be expected to show a too short residence time in the body, and show a lower bioavailability relative to the metabolically stable compounds. The compounds of the present invention have half-lives in HLM of> 110 min.

The test is carried out as follows: 15 Incubations with microsomes

All incubations were performed in a shaken water bath set with a thermostat at 37 ° C. Each incubate contained 0.5 μΜ CYP. Cofactors were added. as a NADPH regeneration system. It consisted of 1.2 mM NADP.5. mM MgCl 2 x 6 H 2 O. 5 mM DL isocitric acid and one unit / ml isocitric acid hydrogenase, which was very pure. All reagents were dissolved in phosphate buffer (50 mM; pH = 7> 4). The substrate concentration was 1 μΜ. The substrates were dissolved from acetonitrile, with the final concentration in acetonitrile in the incubation merge being less than 0.1% (vol / vol). NADP was omitted from the control incubations. In all experiments, the samples were pre-incubated for 5 minutes at 37 ° C with microsomes, substrate and regeneration system, with NADP missing. The reaction was started by adding NADP. The incubation period was 1 hour. Volumes of 100 μΐ were removed after 0, 3, 5, 10, 15, 20, 30,. 45 and 60 minutes. The volumes were extracted with 400 μΐ 1025709

32 I

1M acetic acid and 2.0 ml of ethyl acetate and analyzed by I

by means of LC-MS-MS. I

The compounds of the present invention can be I

be combined with one or more other active compounds

5 things selected from the list: I

1) One or more naturally occurring or synthetic I

prostaglandins or esters thereof. Suitable prostaglan

dines for use herein include compounds such as al-I

prostadil, prostaglandin E1, prostaglandin E0, 13.14-1

Dihydroprostaglandin Ei, prostaglandin Es, eprostiniol, I

natural, synthetic and semi-synthetic prostaglan I

dines and derivatives thereof, including those which I

are described in WO 00033825 and / or US 6,037,346, published

March 14, 2000, all of which are incorporated herein

By reference, PGE0, PGEi, PGAi, PGBi, PGFi a, 19-1

hydroxy-PGA1, 19-hydroxy-PGB1, PGE2 # PGB2, 19-hydroxy-PGA2, I

19-hydroxy-PGB2, PGE3a, carboprosttromethamine, dinoprost-I

tromethamine, dinoprostone, lipoprost, gemeprost, measuring I

oprost, sulprostune, tiaprost and moxisylate. I

2) One or more compounds that are antagonist of the I

α-adrenergic receptor, also known as α-adrenoceptors I

or α-receptors or α-blockers. Suitable compounds I

for use herein include: the α-adrenegic receptor I

blockers, as described in PCT patent application

WO 99/30697, published on June 14, 1998, of which the I

descriptions relating to α-adrenergic re- I

ceptors are incorporated herein by reference, and this I

include selective (Χι-adrenoceptor or α2-I

adrenoceptor blockers and non-selective adrenoceptor blockers, suitable alpha adrenoceptor blockers include:

mine, phentolamine mesylate, trazodone, alfuzosin, indoramine

ne, naftopidil, tamsulosin, dapiprazole, phenoxybenzamine, I

idazoxan, epharaxan, yohimbine, rauwolfia alkaloids, Re-I

cordati 15/2739, SNAP 1069, SNAP 5089, RS17053, SL I

1025709 ___ | 33 89,0591, doxazosin ,. terazosin, abanoquil and prazosin; α 2 -adrenoceptor blockers from US 6,037,346 [March 14, 2000] are dibenamine, tolazoline, trimazosin and dibenamine, α-adrenergic receptors as described in U.S. Patent Nos. 4,188,390; 4,026,894; 3,511,836; 4,315,007; 3,527,761; 3,997,666; 2,503,059; 4,703,063; 3,381,009; 4,252,721 and 2,599,000, all of which are incorporated herein by reference; α 2 -adrenoceptor blockers include: clonidine, papaverine, pa-paverine hydrochloride, optionally in the presence of a cariotonic agent such as pirxamine; 3) One or more NO donor (NO agonist) compounds. Suitable NO donor compounds for use herein include organic nitrates, such as mono-, di- or tri-nitrates, or organic nitrate esters including glyceryl trinitrate (also known as nitroglycerin), isosorbide 5 mononitrate, isosorbide dinitrate, pentaerythritol tetranitrate, erythrityrate tetrate sodium nitroprusside (SNP), 3-morpholino-sydnonimine, molsidomine, S-nitroso-N-acetylpenicillamine (SNAP), S-nitroso-N-glutathione (SNO-GLU), N-hydroxy-L-arginine, amyl nitrate, linsidomine, linsidomine hydrochloride, (SIN-1), S-nitroso-N-cysteine, diazenium diolates, (NONOates), 1,5-pentanedinitrate, L-arginine, ginseng, zizphi fructus, molsidomine, Re-2047, nitrosylated max-25 isylytivates such as NMI-678-11 and NMI-937 as described in published PCT patent application WO 0012075.

4) One or more potassium channel openers or modulators. Suitable potassium channel openers / modulators for use herein include nicorandil, cromocalim, lev-cromakalim, lemakalim, pinacidil, cliazoxide, minoxidil, charybdotoxin, glyburide, 4-aminopyridine, BaCl 2.

5) One or more dopaminergic agents, preferably apomorphine or a selective D-2, D-3 or D2 / D3 agonist such as 1025709

34

Pramipexole and ropirinol (claimed)

I in WO 0023056), PNU 95666 (which is claimed I

I in WO 0040226). I

6) One or more vasodilators. Suitable I

Vasodilators for use herein include Ni

I modepine, pinacidil, cyclandelate, isoxsuprine, chloroprime

Izine, haloperidol, Ree 15/2739, trazodone. I

7) One or more agonists of thromboxane A2. I

I 8) One or more agents that are active on the CNS. I

9) One or more ergot alkaloids. Suitable ergot I

I alkaloids are described in U.S. Patent I

I 6,037,346, published March 14, 2000, and include ace-I

I I

tergamine, brazergoline, bromerguride, cianergoline, del-I

Orgotril, disulergine, ergonovine maleate, ergotamine tare

Iate, etisulergine, lergotril, lysergide, mesulergine, I

I metergoline, metergotamine, nicergoline, pergolide, propyl

I sergide, proterguride, terguride. I

10) One or more compounds that enhance the action of natural I

Modeling of I tical factors, in particular atrial natruretic I

Factor (also known as atrial natruretic peptide), na

I truretic factors of type B and type C, such as inhibitors I

I of neutral endopeptidase, I

11) One or more compounds that convert angiotensin

I inhibiting enzyme inhibitors, such as enapril, and combined inhibitors

I of angiotensin converting enzyme and neutral endopepti

I dase, such as omapatrilat. I

12) One or more angiotensin receptor antagonists, I

I like losartan. I

13) One or more substrates for NO synthase, such as L-I

Arginine. I

14) One or more blockers of the calcium channel, such as am-I

I lodipine. I

15) One or more antagonists of endothelin receptors and I

I inhibitors of endothelin converting enzyme. I

I 1025709 I

16) One or more cholesterol-lowering agents, such as statins (e.g., atorvastatin / Lipitor trademark) and fibrates.

17) One or more platelet anti-caking agents and antithrombotic agents, e.g. tPA, uPA, warfarin, hirudin and other thrombin inhibitors, heparin, thromboplastin activating factor inhibitors.

18) One or more insulin sensitizing agents, such as rezulin and hypoglycaemic agents such as glipizide.

19) L-DOPA or carbidopa.

20) One or more inhibitors of acetylcholinesterase, such as donezipil.

21) One or more steroidal or non-steroidal anti-inflammatory agents.

22) One or more modulators of the estrogen receptor and / or agonists of estrogen and / or antagonists of estrogen, preferably raloxifene, tibolone or lasofoxifene, (-) - cis-6-phenyl-5- [4- (2- pyrrolidin-1-yl) ethoxy) phenyl] 5,6,7,8-tetrahydronaphthalene-2-ol and pharmaceutically acceptable salts thereof, the preparation of which is described in detail in WO 96/21656.

23) One or more modulators of cannabinoid receptors.

24) One or more inhibitors of an NPY (neuropeptide Y), more particularly inhibitors of NPY1 or NPY5, preferably an inhibitor of NPY1, wherein the NPY inhibitors (including NPY Y1. And NPY Y5) at preferably have an IC 50 of less than 100 nM and more preferably of less than 50 nM. An assay for identifying NPY inhibitors is given in WO-A-98/52890 (see pages 96, lines 2 to 28).

25) One or more analogs of vasoactive intestinal protein (VIP), viP mimetics, VIP analogue, which are more specifically intervened by one or more of the VIP receptor subtypes VPAC1, VPAC or PACAP (pituitory adenylate 35 cyclase activating peptide ), one or more of a VIP 1025709

36 I

receptor agonist or a VIP analogue (e.g. Ro-125-1553) or I

a VIP fragment, one or more of an oc

adrenoceptor antagonist with a VIP combination (e.g. Invi- I

corp, Aviptadil). I

26) One or more of a melanocortin receptor agonist or I

a modulator or a melanocortin enhancer, such as melamine

notan II, PT-14, PT-141 or compounds covered by claim I

is made in WO 09964002, WO 00074679, WO 09955679, WO I

00105401, WO 00058361, WO 00114879, WO 00113112, WO I

10 09954358. I

27) One or more of a serotonin receptor agonist, - I

antagonist or modulator, more in particular agonists, I

5HT1A- antagonists or modulators (including VML I

670), 5HT2A, 5HT2C, 5HT3 and / or 5HT6 receptors, including

15 of those described in WO 09902159, WO I

00002550 and / or WO 00028993. I

28) One or more androgens such as androsterone, dehydro-I

androsterone, testosterone, androstanedione and a synthetic

chemical androgen. I

29) One or more estrogens, such as estradiol, estrone, I

estriol and a synthetic estrogen, such as estrogen-I

benzoate. I

30) One or more modulators of carriers for normal

drenaline, dopamine and / or serotonin, such as bupropion, GW-I

25 320659. I

31) One or more agonists and / or modulators of a pu I

rhinergic receptor. I

32) One or more antagonists of a neurokinin (NK) - I

receptor, including those described in WO-I

30 09964008. I

33) One or more agonists, antagonists or modulators I

of an opioid receptor, preferably agonists of the I

ORL-1 receptor. I

1025709 | 34) One or more oxytocin / vasopressin receptor agonists or modulators, preferably an oxytocin selective agonist or modulator.

35) One or more PDE inhibitors, more in particular an inhibitor of PDE 2, 3, 4, 5, 7 or 8, preferably one. inhibitor of PDE2 or PDE5, and most preferably an inhibitor of PDE5 (see below), wherein these inhibitors preferably have an IC 50 value against the respective enzyme of less than 100 nM. Suitable inhibitors of cGMP PDE5 for use in accordance with the present invention include: the pyrazolo [4,3-d] pyrimidine-7-ones described in EP-A-0463756, the pyrazolo [4,3-d] pyrimidine- 7-ones described in EP-A-0526004, the pyrazolo [4,3-d] pyrimidine-7-ones described in published international patent application WO 93/06104, the isomeric pyrazolo [3,4-d] pyrimidine-4-ones described in published international patent application WO 93/07149, the chuinazoline-4-ones described in published international patent application WO 93/12095, the pyrido [3,2-d] pyrimidine-4-ones described in published international patent application WO 94/05661, the purine 6-ones described in published international patent application WO 94/00453, the pyrazolo [4,3-d] pyrimidine-7-ones described in published International Patent Application WO 98/49166, the pyrazolo [4,3-d] pyrimidine-7-ones described In published international patent application WO 99/54333, the pyrazolo [4,3-d] pyrimidine-4-ones described in EP-A-0995751, 1025709

I 38 I

The pyrazolo [4,3-d] pyrimidine-7-ones described I

I in published international patent application WO I

I 00/24745, I

The pyrazolo [4,3-d] pyrimidine-4-ones described I

15 in EP-A-0995750, I

I the compounds described in published I-I

International Patent Application WO 95/19978, I

I the compounds described in published I-I

International Patent Application WO 99/24433, I

The compounds described in published in

International Patent Application WO 93/07124, I

The pyrazolo [4,3-d] pyrimidine-7-ones described I

I in published international patent application WO I

I 01/27112, I

The pyrazolo [4,3-d] pyrimidine-7-ones described I

I in published international patent application WO I

I 01/27113, I

I the compounds described in EP-A-1092718 and I

The compounds described in EP-A-1092719. I

Other suitable inhibitors of PDE5 for use

I according to the present invention include: I

5- [2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) phenyl] -1-I

1-methyl-3-n-propyl-6-dihydro-7 H -pyrazolo [4,3-I

I d] pyrimidine-7-one (sildenafil), also known as 1-t [3- (6,7-I)

Dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo [4,3-I

D) pyrimidin-5-yl) -4-ethoxyphenyl] sulfonyl] -4- I

Methylpiperazine (see EP-A-0463756); I

5- (2-ethoxy-5-morpholinoacetylphenyl) -1-methyl-3-n-propyl-1

1,2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidin-7-one I

30 (see EP-A-0526004); I

3-ethyl-5- [5- (4-ethyl-piperazine-1-ylsulphonyl) -2-n-1

Propoxyphenyl] -2- (pyridin-2-yl) methyl-2,6-dihydro-7 H -1

Pyrazolo [4,3-d] pyrimidin-7-one (see WO 98/49166); I

10257-9 I

39 3-ethyl-5- [5- (4-ethylpiperazine-1-ylsulfonyl) -2- (2-methoxyethoxy) pyridin-3-yl] -2- (pyridin-2-yl) methyl-2,6-dihydro -7 H -pyrazolo [4,3-d] pyrimidin-7-one (see WO 99/54333); 5 (+) - 3-ethyl-5- [5- (4-ethylpiperazine-1-ylsulfonyl) -2- (2-methoxy-1 (R) -methylethoxy) pyridin-3-yl] -2-methyl-2 6-dihydro-7 H -pyrazolo [4,3-d] pyrimidin-7-one, also known as 3-ethyl-5- {5- [4-ethyl piperazine-1-ylsulfonyl] -2 - ([(IR)) -2-methoxy-1-methylethyl] oxy) pyridin-3-yl} -2-methyl-2,6-10 dihydro-7 H -pyrazolo [4,3-d] pyrimidine-7-one (see WO 99/54333 ); 5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfonyl) pyridin-3-yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-7 H -pyrazolo [4.3 -d] pyrimidin-7-one, also known as 1- {6-15 ethoxy-5- [3-ethyl-6,7-dihydro-2- (2-methoxyethyl) -7-oxo-2H-pyrazolo [4 3-d] pyrimidin-5-yl] -3-pyridylsulfonyl} -4-ethylpiperazine (see WO 01/27113, Example 8); 5- [2-iso-butoxy-5- (4-ethyl-piperazin-1-ylsulphonyl) -pyridin-3-yl] -3-ethyl-2- (1-methyl-piperidin-4-20-yl) -2,6-dihydro- 7 H -pyrazolo [4,3-d] pyrimidin-7-one (see WO 01/27133, example 15); 5- [2-ethoxy-5- (4-ethyl-piperazine-1-ylsulfonyl) -pyridin-3-yl] -3-ethyl-2-phenyl-2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine 7-one (see WO 01/27113, example 66); 5- (5-acetyl-2-propoxy-3-pyridinyl) -3-ethyl-2- (1-isopropyl-3-azetidinyl) -2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine -7-one (see WO 01/27112, example 124); 5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine- 7-on 30 (see WO 01/27112, example 132); (6R, 12aR) -2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4-methylenedioxyphenyl) pyrazino [2 ', 1': 6.1] pyrido [3,4 -b] indole-1,4-dione (IC-351), that is, the compound of examples 78 and 95 of published intemio 1025709

I 40 I

International patent application WO 95/19978, as well as the compound I

I of examples 1, 3, 7 and 8; I

2-E2-ethoxy-5- (4-ethyl-piperazin-1-yl-1-sulfonyl) -phenyl] -5-

Methyl 7-propyl-3 H-imidazo [5,1-f] [1,2,4] triazine-4-one

I5 (vardenafil) also known as 1 - [[3- (3,4-dihydro-5-methyl-4-)

Oxo-7-propylimidazo [5,1-f] -as-triazin-2-yl) -4-I

Ethoxyphenyl] sulfonyl] -4-ethylpiperazine, i.e. the I

I compounds of examples 20, 19, 337 and 336 of pu I

International patent application WO 99/24433, and I

I 10 the compound of Example 11 of published international I

International patent application WO 93/07124 (EISAI), and the compound I

Items 3 and 14 of Rotella D.P., J. Med. Chem. 2000, 43, I

I 1257. I

Still other suitable inhibitors of PDE5 include: 4- I

Bromo-5- (pyridylmethylamino) -6- [3- (4-chlorophenyl) propoxy] -1

I 3- (2 H) pyridazinone; I

1- [4 - [(1,3-benzodioxol-5-ylmethyl) amino] -6-chloro-2-quinozolinyl] -4-piperidinecarboxylic acid, monosodium salt;

I (+) - cis-5,6a, 7,9,9a-hexahydro-2- [4- (trifluoromethyl) phenyl]

Methyl 5-methylcyclopent [4,5] imidazo [2,1-b] purine-4 (3H) one; I

Furazlocillin; I

Cis-2-hexyl-5-methyl-3,4,5,6a, 7,8,9,9a-octahydrocyclo-I

I pent [4,5] imidazo [2,1-b] purine-4-one; I

1 3-acetyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylate; I

3-acetyl-1- (2-chlorobenzyl) -2-propylindole-6-carboxylate; I

4-bromo-5- (3-pyridylmethylamino) -6- (3- (4-chlorophenyl) pro-)

Poxy) -3- (2H) pyridazinone; I

1-methyl-5- (5-morpholinoacetyl-2-n-propoxyphenyl) -3-n-pro-

1 pyl-1,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidin-7-one; I

1- [4 - [(1,3-benzodioxol-5-ylmethyl) amino] -6-chloro-2- I

Chuinazolinyl] -4-piperidinecarboxylic acid, sodium salt; I

I Pharmaprojeets No. 4516 (Glaxo Wellcome); I

Pharmaprojeets No. 5051 (Bayer); I

Pharmaprojeets No. 5064 (Kyowa Hakko; see WO 96/26940); I

Pharmaprojeets No. 5069 (Schering Plow); I

1025709 I

41 GF-196960 (Glaxo Wellcome); E-8010 and E-4010 (Eisai);

Bay-38-3045 & 38-9456 (Bayer) and Sch-51866.

5 for it. treatment of FSD, the compounds of the present invention can preferably be combined with one or more active ingredients selected from the list: a) an inhibitor of PDE5, more preferably 5- [2-ethoxy-5- (4-methyl-10). 1-piperazinylsulfonyl) phenyl] -1-methyl-3-n-propyl-1,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine-7-one (sildenafil); (6R, 12aR) -2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4-methylenedioxyphenyl) pyraizino [2 ', 1': 6.1] pyrido [ 3,4-b] indole -1,4-dione (IC-351); 2- [2-Ethoxy-5- (4-ethyl-piperazin-1-yl-1-sulfonyl) -phenyl] -5-methyl-7-propyl-3 H -imidazo [5,1-f] [1,2,4 ] triazine-4-one (vardenafil); 5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfonyl) pyridin-3-yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-7 H -pyrazolo-20 [4 , 3-d] pyrimidin-7-one and 5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2,6-dihydro-7H pyrazolo [4,3-d] pyrimidin-7-one and pharmaceutically acceptable salts thereof; b) an inhibitor of NPY Y1; C) an agonist of dopamine, such as apomorphine or a selective agonist of D2, D3 or D2 / D3, such as pramipexole and ropirinol; d) a melanocortin receptor agonist or modulator, or a melanocortin enhancer, preferably melanotan II, PT-14, PT-141; e) an agonist, antagonist or modulator of 5HT2C; f) a modulator of the estrogen receptor, agonists of estrogen and / or antagonists of estrogen, preferably raloxifene, tibolone or lasofoxifene; 1025709

42 I

g) an androgen, such as androsterone, dehydroandrosterone, I

testosterone, androstanedione and a synthetic androgen I

and I

h) an estrogen, such as estradiol, estrone, estriol I

5 and a synthetic estrogen, such as estrogen benzoate. I

For the treatment of MED, the compounds I

of the present invention are preferably combined

with one or more active ingredients that are produced

chose from the list: I

A) an inhibitor of PDE5, more preferably 5- [2-ethoxy-5- (4-methyl-1)

1-piperazinylsulfonyl) phenyl] -1-methyl-3-n-propyl-1,6-l

dihydro-7 H -pyrazolo [4,3-d] pyrimidin-7-one (sildenafil); I

(6R, 12aR) -2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4-I

methylenedioxyphenyl) pyrazino [2 ', 1': 6,1] pyrido [3,4-I

B) indole-1,4-dione (IC-351); I

2- [2-ethoxy-5- (4-ethylpiperazin-1-yl-1-sulfonyl) phenyl] -5-I

methyl 7-propyl-3 H -imidazo [5,1-f] [1,2,4] triazine-4-one I

(vardenafil); I

5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfonyl) pyridine-3-I

20 yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-7 H -pyrazolo

[4,3-d] pyrimidin-7-one and I

5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-)

azetidinyl) -2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine-7-one I

and pharmaceutically acceptable salts thereof; I

B) an inhibitor of NPY Y1; I

c) an dopamine agonist (preferably apomorphine) or I

a selective agonist of D2, D3 or D2 / D3, such as prami

pexole and ropirinol; I

d) an agonist or modulator of the melanocortin I

Receptor, or an melanocortin enhancer, for example

prefer melanotan II, PT-14, PT-141 and I

e) an agonist, antagonist or modulator of 5HT2C. I

Combinations used to treat FSD in particular I

preferred are the compounds of I below

1025709 I

The present invention and one or more active ingredients selected from the list are 5- [2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) phenyl] -1-methyl-3-n-propyl-1,6 dihydro-7 H -pyrazolo [4,3-d] pyrimi-5-din-7-one (sildenafil); (6R, 12aR) -2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4-methylenedioxyphenyl) pyrazino [2 ', 1': 6.1] pyrido [3,4 -b] indole-1,4-dione (IC-351); 2- [2-ethoxy-5- (4-ethyl-piperazin-1-yl-1-sulfonyl) -phenyl] -5-10-methyl-7-propyl-3 H -imidazo [5,1-f] [1,2,4 ] triazine-4-one (vardenafil); 5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfonyl) pyridin-3-yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-7 H -pyrazolo [4, 3-d] pyrimidin-7-one and 5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2,6-dihydro-7H pyrazolo [4,3-d] pyrimidin-7-one; apomorphine; melanotan II ;.

PT-141; Lasofoxifene; raloxifene; tibolone; an androgen, such as androsterone, dehydroandrosterone, testosterone, androstanedione and a synthetic androgen, and an estrogen, such as estradiol, estrone, estriol, and a synthetic estrogen such as estrogen benzoate.

Particularly preferred combinations for treating MED are the compounds of the present invention and one or more active ingredients selected from the list: 5- [2-ethoxy-5- (4-methyl-1-) piperazinylsulfonyl) phenyl] -1-methyl-3-n-propyl-1,6-dihydro-7 H -pyrazolo [4,3-d] pyrimi. dine-7-one (sildenafil); 1025709

44 I

(6R, 12aR) -2,3,6,7,12,12a-hexahydro-2-methyl-6- (3,4-methyl)

lendioxyphenyl) pyrazino [2 ', 1': 6,1] pyrido [3,4-b] indole-1,4

dione (IC-351); I

2 - [[2-ethoxy-5- (4-ethylpiperazin-1-yl-1-sulfonyl) phenyl] -5-I

5-methyl-7-propyl-3 H -imidazo [5,1-f] [1,2,4] triazine-4-qn I

(vardenafil); I

5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfonyl) pyridine-3-I

yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-7 H -pyrazolo

[4,3-d] pyrimidin-7-one and I

5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-)

azetidinyl) -2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidin-7-one; I

apomorphine; I

melanotan II and I

PT-141. I

15 For the treatment of cardiovascular disorders, I

in particular hypertension, the compounds of I

the present invention can be combined with one or I

more active ingredients selected from the list I

m: I

A) blockers of the angiotensin receptor (ARB), such as I

losartan, valsartan, telmisartan, candesartan, irbesartan, I

eprosartan and olmesartan; I

b) calcium channel blockers (CCB), such as amlodipi

ne; I

C) statins, such as atorvastatin; I

d) inhibitors of PDE5, such as sildenafyl, tadalafil, var

denafil, 5- [2-ethoxy-5- (4-ethylpiperazine-1-ylsulfol)

methyl) pyridin-3-yl] -3-ethyl-2- [2-methoxyethyl] -2,6-dihydro-1

7 H -pyrazolo [4,3-d] pyrimidin-7-one; I

5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-)

azetidinyl) -2,6-dihydro-7 H -pyrazolo [4,3-d] pyrimidine-7-one I

and I

the pyrazolo [4,3-d] pyrimidine-4-ones described I

in WO 00/27848, in particular N - [[3 - (4,7-dihydro-1-)

Methyl 7-oxo-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl) -1

1025709 I

45 4-propoxyphenyl] sulfonyl] -1-methyl-2-pyrrolidine propanamide [DA-8159 (Example 68 of WO 00/27848)]; e) beta blockers, such as atenolol or carvedilol; f) ACE inhibitors such as quinapril, enelapril and lisinopril; g) alpha blockers such as doxazosin; h) Aldosterone receptor selective antagonists (SARA), such as eplerenone or spironolactone, and i) Imidazoline II agonists such as rilmenidine and moxonidine.

If a combination of active agents is administered, these can be administered simultaneously, separately or sequentially.

The compounds of the present invention can be administered alone, but in human therapy this will generally be done in admixture with a suitable pharmaceutical excipient, diluent, or carrier selected for the intended route of administration. and the standard pharmaceutical practice.

The present invention provides a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts, solvates or polymorphs thereof, and a pharmaceutically acceptable diluent or carrier.

Oral administration

The compounds of the present invention can be administered orally. Oral administration may involve ingestion, so that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be used, wherein the compound enters the blood stream directly through the mouth.

Formulations suitable for oral administration include solid formulations, such as tablets, capsules 1025709

46 I

contain solid particles, liquids or powders, pastil I

lesson (including liquid-filled), chewing gums, prepa I

combs that consist of very many small or nanoparticles, I

gels, films (including mucoadhesive), ovules, sprays and I

Liquid preparations. I

Liquid preparations include suspensions, solutions

genes, syrups and elixirs. Such compositions may be I

are used as fillers in soft or hard capsules

lesson and typically include a carrier, e.g.

10 water, ethanol, propylene glycol, methyl cellulose, or I

a suitable oil, and one or more emulsifiers and / or suspension

spending means. Liquid preparations can also be I

prepared by the renewed composition of a fixed I

substance, for example from a sachet. I

The compounds of the present invention can be I

can also be used in fast dissolving, fast disintegration I

different dosage forms such as those described in I

Expert Opinion in Therapeutic Patents, 11 (6), 981-986, I

by Liang and Chen (2001). I

A typical tablet can be prepared using I

standard methods used for a formulation chemist

are known, for example by direct compression, I

granulation (dry, wet, or melting), melting and solidifying, I

or extrusion. The tablet preparation may be from one or more of the above

25 exist and whether or not they are lined. I

Examples of excipients suitable for I

oral administration include carriers, for example cellulose, I

calcium carbonate, dibasic calcium phosphate, mannitol and I

sodium citrate, binders for granulation, e.g.

Polyvinylpyrrolidine, hydroxypropylcellulose, hydroxy

hydroxypropyl methylcellulose and gelatin, disintegration

gelling agents, for example sodium starch glycolate and silica

licenses, lubricants, for example magnesium stearate and I

stearic acid, wetting agents, for example sodium I

1025709 I

47 lauryl sulfate, preservatives, ainti-oxidants, flavors and colorings.

Solid compositions for oral administration can be formulated for immediate and / or modified release. Modified-release preparations include delayed, continuous, pulsed, controlled double, targeted, and programmed release. . The details of suitable modified release technologies, such as high-energy dispersions, osmotic and coated particles can be found in Pharmaceutical Technology On-line, Verma et al., 25 (2), 1-14 (2001). Other modified release preparations are described in U.S. Patent No. 6,106,864.

.15

Parenteral administration

The compounds of the present invention can also be administered directly into the blood stream, into the muscles or into an internal organ. Suitable modes of parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intrauretral, intrasternal, intracranial, intramuscular. and subcutaneous administration. Suitable means of parenteral administration include needle (including micro needle) injectors, needle free injectors and infusion techniques.

Parenteral compositions are typically aqueous solutions containing excipients such as salts, carbohydrates, and buffering agents (preferably up to a pH of 30 to 9), but for some applications, they may suitably be formulated as a sterile, non-aqueous solution or as a dried form that will be used with a suitable vehicle such as sterile, pyrogen-free water.

1025709

48 I

The preparation of parenteral preparations under sterile I

Under certain circumstances, for example by freeze drying, I

easily achieved with the help of pharmaceuticals

standard technical techniques known to those skilled in the art. I

5 I

The solubility of compounds of formula (I) that I

in the preparation of parenteral solutions, I

can be increased by a suitable operation, I

for example through the use of high-energy gel

Spray-dried dispersions (see WO 01/47495) and / or through the I

use of the correct formulation techniques, such as by I

the use of agents that increase solubility. I

Preparations for parenteral administration can be I

formulated for immediate and / or modified I

15 donation. Modified-release preparations include I

those for delayed, uninterrupted, pulsed, controlled I

learned dual, targeted and programmed release

to. I

Topical administration I

The compounds of the present invention can be I

also be topically applied to the skin (preferably on I

the genitals) or mucous membranes, both dermal and trans-I

dermal. Typical compositions for this purpose include gel

25, hydrogels, lotions, solutions, creams, ointments, I

dusting powders, spreads, foams, films, skin patches, I

waffles, implants, sponges, fibers, dressings and micro-I

emulsions. Liposomes can also be used. Typical I

carriers include alcohol, water, mineral oil, liquid I

Petrolatum, white petrolatum, glycerin and propylene glycol

col Penetration enhancers can be included (see I

for example Finnin and Morgan, J. Pharm. Sci., 88 (10), I

955-958 (October 1999). I

Other modes of topical administration include delivery

35 by iontophoresis, electroporation, phonophoresis, sonophoresis

1025700 I

49 s and needle-free injection or injection using micronaids.

Preparations for topical administration can be formulated for immediate and / or modified release. Modified-release preparations include delayed, continuous, pulsed, controlled, dual, targeted, and programmed release preparations. Thus, the compounds of the present invention can be formulated for administration in a more solid form, an implanted depot preparation, which provides for a sustained release of the active compound.

Inhaled / intranasal administration

The compounds of the present invention can also be administered intranasally or by inhalation, typically in the form of a dry powder (either alone, as a mixture, for example in a dry mixture with lactose, or as a preparation of mixed particles , for example mixed with phospholipids) from a dry powder inhaler, or as an aerosol spray from a pressurized container, pump, spray, nebulizer (preferably a nebulizer that produces a fine mist with the help of electrohydrodynamics), with or without the use of a suitable propellant such as dichlorofluoromethane.

The pressurized container, pump, spray or atomizer contains a solution or suspension of the active compound which, for example, ethanol (optionally aqueous ethanol) or a suitable alternative means for dispersing, solubilizing or assisting in the delivery of 30 the active agent, propellant (s) as solvent and an optional surfactant, such as sorbitan triolate or an oligamic acid.

Prior to use in a dry powder or suspension preparation, the drug product is reduced to a size suitable for delivery by inhalation. 102.5 70 9

50 I

(typically less than 5 microns). This can be I

be realized by means of a suitable reduction

process, such as grinding by means of a screw

jet, grinding by means of a fluid bed jet, working I

5 with a supercritical fluid, wherein nanoparticles

particles are formed, high pressure homogenization or I

spray drying. I

A suitable preparation in the form of a solution I

for use in a nebuliser that uses electric I

10 trohydrodynamics produces a fine mist, 1 μg to I

10 mg of the compound of the present invention per I

stroke, and the stroke volume can range from 1 μΐ I

up to 100 μΐ. A typical preparation may be a compound with I

formula (I), propylene glycol, sterile water, ethanol and I

Contain sodium chloride. Alternative solvents that I

can be used instead of propylene glycol, om

glycerol and polyethylene glycol. I

Capsules, blister packs and cartridges (which I

made from gelatin or HPMC) for use I

20 in an inhaler or insufflator can be so applied

formulated that it is a powder mixture of the compound I

of the present invention, a suitable powder base

substance such as lactose or starch, and a performance modifier I

such as L-leucine, mannitol or magnesium stearate. I

With dry powder inhalers and aerosols, the do-I

unit determined by means of a valve that has an I

measured amount. The units in accordance with I

in accordance with the present invention are in a type I

case is set so that a measured dose or "puff" I

30 is administered containing 1 μg to 50 mg of the compound with I

the formula (I). The total daily dose will be in I

a typical case in the range of 1 mg to 50 mg, such as I

1 mg to 50 mg, and this can be in a single dose

1025709 I

51 sis, or more often, if divided doses are administered during the day.

Preparations for inhaled / intranasal administration can be formulated for immediate and / or modified release. Modified, sustained release preparations include delayed, continuous, pulsed, controlled, dual, targeted, and programmed release preparations.

10 Rectal / intravaginal administration

The compounds of the present invention can be administered rectally or vaginally, for example in the form of a suppository, pessary or clysma. Cocoa butter is a traditional raw material for a suppository, but depending on what is applicable, various alternatives can be used. Preparations for rectal / vaginal administration may be formulated for immediate and / or modified release. Preparations for modified release include preparations for the delayed, continuous, pulsed, controlled bipartisan, targeted and programmed release.

Ocular / andial administration.

The compounds of the present invention can also be administered directly to the eye or ear, typically in the form of drops of a finely divided suspension or solution in isotonic, sterile saline whose pH is controlled. Other compositions suitable for ocular and andial administration include ointments, biodegradable (e.g. absorbable gel sponges, collagen) and non-biodegradable (e.g. silicone) implants, waffles, lenses and solid particle or vesicle systems, such as niosomes or liposomes. A polymer such as a cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, a cellulose poly 1025709

52 I

more, for example hydroxypropyl methyl cellulose, hydroxy I

ethylcellulose or methylcellulose or a heteropolysaccha I

red polymer, for example, as appropriate, can be included I

together with a preservative such as benzalkonium chloride

5 ride. Such compositions may also be delivered

by iontophoresis. I

Preparations for ocular / andial administration may I

be formulated for immediate and / or modified I

the issue. Modified-release preparations include I

10 preparations for the delayed, continuous, pulsed I

controlled, controlled dual, targeted or pro- I

programmed release. I

Required technologies I

The compounds of the present invention can be I

be combined with soluble macromolecular entity

such as cyclodextrin or polymers containing polyethylene I

glycol for the purpose of improving their opacity

releasability, dissolution rate, masking the I

Taste, bioavailability and / or stability. I

Drug / cyclodextrin complexes are for example

example has generally proved useful for most I

dosage forms and administration routes. Both inclusion and I

non-inclusion complexes can be used. If an I

25 alternative to direct complexation with the gel

the drug, the cyclodextrin can be used as an auxiliary additive

used as a carrier, diluent or soap

lubilizer. For these purposes, they are most often used

fa, beta and gamma cyclodextrins are used, of which before I

30 images can be found in International oc

WO 91/11172, WO 94/02518 and WO 98/55148. I

Dosage I

For administration to human patients, the toll is I

35 daily dose of the compounds of the present invention

1025709 I

The invention is typically between 0.1 mg and 1000 mg, which, of course, depends on the method of administration. For example, oral administration may require a total daily dose of 5 mg to 1000 mg, such as 5 to 500 mg, while an intravenous dose requires only 0.01 to 30 mg / kg of body weight, such as 0.1 to 10 mg / kg, more preferably May require 0.1 to 1 mg / kg body weight. The total daily dose can be administered in single or divided doses.

These dosages are based on an average human person weighing about 65 to 70 kg. The doctor will be able to determine doses immediately for persons whose weight falls outside this range, such as children and the elderly.

Those skilled in the art will also appreciate that in the treatment of certain conditions (including FSD and MED), the compounds of the present invention may be taken as a single dose based on "as needed" (i.e., as needed or desired) .

In a preferred embodiment, the compounds of the present invention are delivered systemically (such as orally, buccally and sublingually) and preferably orally. Such a systemic (preferably oral) administration is preferably used to treat female sexual dysfunction, preferably FSAD.

Thus, in a particularly preferred embodiment, use is made of the compounds of the present invention in the manufacture of a systemically delivered (preferably orally delivered) drug for the treatment or prophylaxis of FSD, preferably FSAD.

A preferred oral preparation uses immediate release or rapidly dispersing or dissolving dosage preparation tablets.

I · 54 I

Fast dispersing or dissolving dosage / formulations; I

I FDDFs). I

In another preferred embodiment, the ver I

I compounds of the present invention have been topically added

Iend, preferably directly to the female genius

I taliën, in particular the vagina. I

I If NEP is present in the entire body, I

I strongly disagree with the expectation that compounds of the I

The present invention can be administered systemically I

10 and a therapeutic response in the female genitalia

I realize without unbearable (harmful) side effects I

I bring about. In EP 1,097,719-A1 and the animal model I

I hereafter we have shown that NEP inhibitors contained in a mo I

For rabbits (in vivo) the I

Blood flow in the genitals after sexual stimulation

I (simulated by stimulating the pelvic nerve) increase I

Without adversely affecting the cardiovascular parameters

I floods, such as by causing a substantial I

I hypotension or hypertension. I

The compounds of the present invention become I

I preferably administered for the treatment of FSD in the I

I sexually stimulated patient (with sexual stimulation I

I refer to audio, visual or touch stimulation). The I

I stimulation may be before, after or during the administration

I 25 den. I

The compounds of the present invention are I

I therefore strengthen the routes / mechanisms on which I am based

I to sexual arousal in the female genitalia, and I

I restore or improve the sexual arousal response in I

I 30 sexual stimulation. I

Thus, a preferred embodiment for the invention is in the case

Use of a compound of the present invention in I

I the preparation of a medicine for treatment or I

I prophylaxis of FSD in the stimulated patient. I

I 1025709 I

55

For veterinary use, a compound of the present invention is administered as a suitable, acceptable preparation in accordance with normal veterinary practice and the veterinarian will determine the dosing regime and route of administration that are most suitable for a particular animal.

The following preparation examples are purely illustrative and are not intended to be used. limit the scope of the present invention. "Active ingredient" means a compound of the present invention.

Preparation 1: a preparation is prepared using the following ingredients: weight / mg

Active ingredient 250

Cellulose, microcrystalline 400

Silicon dioxide, annealed 10

Stearic acid 5

Total 665 The ingredients are mixed and compressed into tablets.

Preparation 2: an intravenous preparation can be prepared as follows:

Active component. 100 mg

Isotonic saline 1000 ml 20

Typical compositions useful for the topical administration of the compounds of the present invention to the genitals are as follows:

Preparation 3: a spray.

Active ingredient (1.0%) in isopropanol (30%) and water.

1025709

56 I

Preparation 4: a foam: I

Active ingredient, glacial acetic acid, benzoic acid, cetyl alcohol

hollow, methyl parahydroxybenzoate, phosphoric acid, polyvinyl alcohol

hollow, propylene glycol, sodium carboxymethyl cellulose, stea I

Rhinic acid, diethyl stearamide, Van Dyke perfume No. 63.01, ge

purified water and isobutane. I

Preparation 5: a gel. I

Active ingredient, docusate sodium BP, isopropylal-I

Alcohol BP, propylene glycol, sodium hydroxide, carbomer I

934P, benzoic acid and purified water. I

Preparation 6: a cream: I

Active ingredient, benzoic acid, cetyl alcohol, long-l

15 del, compound 13091, methyl paraben, propyl paraben, pro I

pylene glycol, sodium carboxymethyl cellulose, sodium I

lauryl sulfate, stearic acid, triethanolamine, glacial acetic acid, I

castor oil, potassium hydroxide, sorbic acid and purified water

ter. I

20 I

Preparation 7: a pessary: I

Active ingredient, cetomacrogol 1000 BP, lemon I

acid, PEG 1500 and 1000 and purified water. I

The present invention furthermore comprises: I

(i) a pharmaceutical composition comprising a compound I

of the present invention, together with a pharmaceutical I

acceptably acceptable excipient, diluent or I

carrier, I

(Ii) a compound of the present invention or I

a pharmaceutically acceptable salt, solvate or polymorph I

thereof, for use as a medicine. I

(iii) the use of a connection from the subject I

the invention as a medicament for treating or I

Prevention of a condition for which a favorable therapy I

1025708 I

A pathological response can be obtained by the inhibition of neutral endopeptidase.

(iv) the use of a compound of the present invention as a medicament for treating or preventing hypoactive sexual appetite disorder, sexual arousal disorder, orgasmic disorder or sexual pain disorder, and preferably sexual arousal disorder, orgasmic disorder or sexual pain disorder, and more preferably sexual arousal disorder.

10 (v) a method for treating PSD or MED

in a mammal, including treating the mammal with an effective amount of a compound of the present invention.

(vi) a pharmaceutical preparation for treating FSD or MED, which comprises a compound of the present invention together with a pharmaceutically acceptable excipient, diluent or carrier.

(vii) a compound of the present invention for use in treating FSD or MED.

(Viii) the use of a compound of the present invention in the manufacture of a medicament for treating or preventing FSD or MED.

The present invention is illustrated by the following non-limiting examples, in which the following abbreviations and definitions are used:

Arbocel® filtering agent br. breed brm breed multiplet

Boe tert-butoxycarbonyl 30 CDI 1,1'-carbonyldiimidazole δ chemical shift d doublet Δ heat DCCI dicyclohexylcarbodiimide 35 DCM dichloromethane 1025709

58 I

DMA dimethylacetamide I

DMF Ν, Ν-dimethylformamide I

DMSO dimethyl sulfoxide I

ES + electrospray ionization, positive scan

5 ES electrospray ionization, negative scan I

Example I

u hours I

HOBt 1-hydroxybenzotriazole I

HPLC high pressure liquid chromatography I

10 m multiplet I

m / z peak in mass spectrum

min. minutes I

m.p. melting point I

MiBK methyl isobutyl ketone I

MS mass spectrum I

NMR nuclear magnetic resonance I

Pree precursor I

Ber. preparation I

k quartet I

20 kwin. quintet I

s singlet I

t triplet I

t-BMA tert-butyl methyl ether I

Tf trifluoromethanesulfonyl I

TFA trifluoroacetic acid I

THF tetrahydrofuran I

TLC thin layer chromatography I

TS + thermospray ionization, positive scan I

WSCDI 1- (3-dimethylaminopropyl) -3-I

Ethylcarbodiimide hydrochloride, in water I

soluble. I

1 H nuclear magnetic resonance (NMR) spectra occurred in all

correspond to the proposed structures. Features- I

the chemical shifts (δ) are given in parts

35 per million from tetramethylsilane, with the initial I

1025709 I

The following abbreviations are used to indicate the main peaks of the usual abbreviations: (e.g. s, singlet; d, doublet; t, triplet; k, quartet; m, multiplet; br, broad. Cl3 deuterochloroform; DMSO, dimethyl sulfoxide. The abbreviation psi means pounds per square inch and LRMS means low-resolution mass spectrometry. Rf is the distance traveled by a connection divided by the distance traveled by the liquid front on a TLC plate The melting points were determined using a Perkin-Elmer DSC7 operating at a heating rate of 20 ° C / minute.

15

Example 1 (R) -2-Methyl-3- (1 - ([3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl) cyclopentyl) propanoic acid 20 DEG C. Tert-Butyl- (2R) -2-methyl-3- [1 - ({[3- (2-methyl-1,3-benzo-thiazol-6-yl) propyl] aminocarbonyl) cyclopentyl] propanoate (preparation 7 and Preparation 8) (7.4 g> 16.7 mmol) was dissolved in dichloromethane (10 ml), trifluoroacetic acid (10 ml) added and the mixture stirred at room temperature for 5 hours. of potassium carbonate (10% aqueous solution) to adjust the pH to about 3 (about 120 ml was needed) The resulting mixture was extracted with 1025709

60 I

dichloromethane (3 x 100 ml), the combined organic I

layers were dried with MgSO * and evaporated. The residue I

was purified by flash chromatography [SiO 2; methanol in I

dichloromethane 1% to 2%], representing the desired acid as I

5 gave a clear oil (5.66 g, 87%). This party I

was combined with 1.4 g of material from a previous I

reaction and stirred in pent on (100 ml) for 3 hours. . I

The pentane layer was removed, the residue scratched about it

loosening gummy residue and for another 2 hours I

Stirred with another portion of pentane (100 ml). The design I

white powder was collected on a glass filter and supported

dried under vacuum at 45 ° C to give the title compound I

as a free-flowing white powder (m.p. 105-1)

106 ° C) (6.52 g). Found: C, 64.93; H, 7.29; N, 7.18. I

C 21 H 28 N 2 O 3 S requires: C, 64.92; H, 7.26; N, 7.21. I

1 H NMR (CDCl 3, 500 MHz) 8 h: 7.85 (1 H, d), 7.61 (1 H, d), 1

7.23 (1H, dd), 5.89 (1H, brm), 3.25-3.35 (2H, m), 2.81 I

(3 H, s), 2.74 (2 H, m), 2.45 (1 H, m), 2.10 (1 H, m), 1.98 I

(1 H, m), 1.89 (1 H, m), 1.88 (2 H, m), 1.55-1.68 (5 H, m), I

20 1.49 (2H, m), 1.17 (3H, d), I

13 C NMR (CDCl 3, 125 MHz) δ0: 180.5, 177.4, 166.7, 151.7, I

138.5, 135.9, 126.7, 122.1, 120.7, 54.5, 42.7, 39.6, 37.4,

36.6, 36.1, 33.4, 31.3, 24.0, 24.3, 19.9, 19.3. I

m / z (electrospray negative ion); 387 (M-H +) I

Measurements of the optical rotation were made in an I

methanol solution (5.7 mg, in 5 ml) with the following I

results: I

[cc] 25S89 -4.4 I

25578 "4.6 I

[a] 2S54e "^ TTi I

- 7.9 l

~~ ta] 25365 I

1025709 I

61

The purity was determined to be> 99% by HPLC analysis with five different inverted phase columns:

Percentage Purity 225 sister 254 am

Main Important - Main Important - Retention peak ste verent peak ate unplanned from cleaning cleaning main peak.min.

Phenomenex Phenyl 100% = 99.95% 0.05% JTS

Hexyl 3 | im

Phenomenex Synergi 99.95% 0.05% 99.9% 0.04%. 3.8

Polar RP 4 | un

Develosil Combi 10% - 99.9% 0.04% 3.9 RP C1 + 3pm

Dionex Acclaim 99.95% 0.05% 99.9% 0.03%. 3.9

Cis 3μπ »

Gravity 99.4% 0.06% 99.9% 0.04% ITS

Cu 3μιη 5 HPLC conditions: Analytical

Temperature: environment

Detection: 225, 254 nm

Mobile phase: A: water: MeCN: TFA

10 95: 5: 0.1% (full / full)

B: MeCN

elution with linear gradient (see below) Flow rate: 1 ml / min.

Conditions solvent gradient: 1025709

62 I

Γ “Time% ~ Β I

0.0 Ö I

_ I

5.0 95 I

ΤΠ 95 I

__ _

The chiral purity was determined to be 98% by I

capillary electrophoresis. This was compared with I

an authentic sample of the opposite enantiomer I

prepared along a similar route, using I

of the conditions described below. I

1025709 I

63 CE conditions

Capillary Agilent, calcined silica, capillary with extended light path of 64.5 cm (effective length 56 cm), 50 μηι I.D.

Temperature 15 ° C

UV detection at 230, 254 and 260 nm

Sample solution approx. 1 mg / ml in running buffer: water: methanol: acetone (1: 10: 1: 0.5)

HP 3DCE system / data file (see attached prints)

Injection 4 seconds of 50 mbar for sample, then 2 seconds of 50 mbar for water Running buffer 250 mg α-cyclodextrin and 50 mg SBE-β-

cyclodextrin, dissolved in borax buffer with pH = 9.3, 50 mM (Agilent CE solution), 5 ml Preparation New capillary: 10 minutes 930 mBar, 1.0 M NaOH

Between measurements: 2 minutes 930 mBar, 0.1 M NaOH, rinse for 2 seconds. with water, 4 minutes with running buffer at 930 mBar

Voltage 25 kV (increasing from 0-25 kV in 30 seconds)

Duration 20 minutes.

An alternative procedure for the production of the title compound is given below: tert-Butyl (2R) -2-methyl-3- [1 - ({[3- (2-methyl-1,3,3-benzothiazole-6-) yl) propyl] amino} carbonyl) cyciopentyl] propanoate (Preparation 7 and Preparation 8) (2.4 g, 5.4 mmol) was dissolved in toluene (7 inl), trifluoroacetic acid (4.1 ml) added and the mixture stirred at 17 ° C for 6 hours. The reaction was terminated by the addition of sodium carbonate 1025709

64 I

bonate (9% solution in water) to adjust the pH to 3 I

(30 ml was needed). MiBK (15 ml) was added. The I

organic phase was separated and the product extracted

in sodium carbonate (9% aqueous solution, 2x5 l

5 ml). The product was extracted in isopropyl acetate I

(35 ml) by adjusting the pH to 4.5 with 5M HCl in 1 hour. I

The organic phase was distilled off under I

atmospheric pressure concentrated to a volume up to 1

of the starting material of 5 ml / g. The I

Cool the oil to ambient temperature, crystallize I

and reduce to granules for 1 hour at 0 to -5 ° C. I

The solid was collected by vacuum filtration, washed with isopropyl acetate (5 ml) and under vacuum

dried at 40 ° C overnight, giving the title

Yielded binding as a free-flowing, white powder (m.p. I

105-106 ° C) [1.3 g, 3.3 mmol, (62%)]. I

Example 2 I

3- (1 - ([3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbam)

20 moyl) cyclopentyl) propanoic acid I

hrr. »- v I

V I

25 O I

This compound was prepared by a procedure

that was analogous to that described in I

Example 1, starting from tert-butyl-3- [1 - ({[3- (2- I

Ethyl 1,3-benzothiazol-6-yl) propyl] aminocarbonyl) cycloyl

pentyl] propanoate from preparation 9. I

M.p. Mp 127.5-129.5 ° C; I

Δ-NMR (d 6 -DMSO, 400 MHz) δ b: 7.89 (1 H, d), 7.63 (1 H, s), 1

7.25 (1H, d), 5.69 (1H, brm), 3.37 (2H, m), 3.15 (2H, k), I

35 2.76 (2H, m), 2.31 (2H, m), 1.96-1.87 (6H, m), 1.70-1.55 I

1025709 I

H

65 (4H, m), 1.47 (3H, t), m / z (ES +): 411 (MNa +), 389 (MH +); m / z (ES "): 387 (M-H +).

Found: C, 64.74; H, 7.28; N, 7.14.

C 21 H 28 N 2 O 3 S requires: C, 64.92; H, 7.26; N, 7.21.

5

The purity was determined to be> 98% by HPLC analysis with three different inverted phase columns:

Percentage Purity 225 nm "" 254 nm ~~~~~~~~

Main Important- Main Important- Retention peak ete most suspected cleaning time cleaning main peak.min.

Luna Phenyl Hexyl 3 99.25% 0.51% 98.79% 0.56% 9.80 μπ \ (150 x 4.6 mm)

Phenoraenex Synergi '99.25% 0.49% 99.14% 0.44% 9.98

Polar RP 4 μιη (150 x 4.6 mm)

Curosil PFP 1 99.31% Ö7Ï% 98.21% 0.84% 9.05 (150 x 4.6 mm) 10 HPLC conditions: Analytical

Temperature: environment

Detection: 225, 254 nm

Mobile phase: A: water: MeCN: TFA

95: 5: 0.1% (full / full)

B: MeCN

elution with linear gradient (see below) Flow rate: 1 ml / min.

Conditions solvent gradient: 1025709 '

66 I

Time (min.)% B I

0.2-15 0-100 l

15-18: ÏÖÖ I

18-18.2 1 100-0

_____ -

Example 3 I

3- (1 - ([3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl-3-carbyl]

5 moyl (Vcyclohexyl) propanoic acid I

"V ^ Y ^ QJ-λ I

0 o I

This compound was prepared by a procedure

15 which was analogous to that described in I

Example 1, starting from tert-butyl-3- (1- {[3- (2- I

ethyl -1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) -1

propanoate from preparation 12. I

^ -NMR (de-DMSO, 400 MHz) ba: 7.86 (1H, d), 7.64 (1H, d), 1

20 7.24 (1H, dd), 5.72 (1H, brs), 3.32 (2H, m), 3.13 (2H, k), I

2.75 (2H, m), 2.27 2H, m), 1.88 {2H, quin), 1.85-1.73 (4H, I)

m), 1.60-1.23 (8 H, m), 1.46 (3 H, t); m / z (ES +): 425 (MNa +), I

403 (MH +); m / z (ES): 401 (M-H +). I

Example 4 I

(R) -2-Methyl-3- (1 - ([3- (2-ethyl-1,3-benzothiazol-6-yl) -1

propyl] carbamoyl) cyclopentyl) propanoic acid I

1025709 I

67 o o 5 ·

The title compound was prepared using the procedure described in Example 1, starting from tert-butyl- (2R) -2-methyl-3- [1 - ({[3- (2-ethyl-1)] 1,3-benzothiazol-6-yl) propyl] amino} carbonyl) cyclopentyl] -10 propanoate from preparation 10.

^ -NMR {CDCl3, 400 MHz) Sgz 7.86 (1H, d), 7.62 (1H, d), 7.24 (1H, dd), 5.81 (1H, brm), 3.27 ( 2H, m), 3.12 (2H, k), 2.74 (2H, m), 2.43 (1H, tn), 2.11-2.05 (1H, tn), 1.98-1 81 (4H, m), 1.69-1.40 (7H, m), 1.45 (3H, t), 1.16 (3H, d); 15 m / z (ES +): 425 (MNa +), 403 (MH +); m / z (ES "): 401 (M-H +).

Example 5 3- (1 - ([3- (2-Methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl) cyclohexyl) propanoic acid 20 h 25%.

The title compound was prepared by a procedure analogous to that described in Example 1, starting from tert-butyl 3- (1- {[3- (2-30 methyl-1,3-benzothiazole)] -6-yl) propyl] carbomoyl} cyclohexyl) propanoate from preparation 13.

^ -NMR (CD3OD, 400 MHz) δκ: 7.78 (1H, d), 7.71 (1H, brm), 7.34 (1H, d), 3.29-3.23 (hidden) ( 2H, m), 2.80 (3H, s), 2.77 (2H, tn), 2.22-2.17 (2H, m), 2.05-1.98 (2H, m), 1 , 92-1025709

68

I 1.85 (2H, m), 1.78-1.74 (2H, m), 1.63-1.51 (3H, m), 1.44-1

I 1.22 (5H, m); m / z (APCI +): 389 (ΜΗ *) I

I Found: I

I C, 64.05; H, 7.39; N, 8.82. I

15 for C 21 H 28 N 2 O 3 S. 0.25 H 2 O is required: I

I C, 64.18; H, 7.31; N, 7.13. I

Example 6

3- (1 - {[3- (2-Methyl-1,3-benzothiazol-6-yl) propyl] carbam

10 moyl) cyclopentyl) propanoic acid I

Ia C3 H - I

I 15 0 0

I I

The title compound was prepared using an I

Procedure analogous to that described in I

Example 1, starting from tert-butyl 3- (1 - {[3- (2- I

Methyl-1,3-benzothiazol-6-yl) propyl] carbomoylcyclo-l

Pentyl) propanoate from preparation 14. I

1 H NMR (CDCl 3, 400 MHz) 6h: 7.85 (1H, d), 7.61 (1H, d), 1

I 25 7.24 (1H, dd), 5.70 (1H, brs), 3.30 (1H, m), 2.81 (3H, s), I

I 2.74 (2H, m), 2.30 (2H, m), 1.97-1.84 (6H, m), 1.69-1.56 I

I (4H, m), 1.50-1.41 (2H, m), - m / z (APCl +): 375 (MH +); (APCI 'I

I): 373 (ΜΗ *). I

Preparation 30 I

Pi (tert-butyl) -3- (2-methyl-1,3-benzothiazol-6-yl) pro- I

Pyramidodicarbonate I

I 1025709_

35 I

69 '^

. ° Y ° jfTV

.5 -, τ °;

Di (tert-butyl) allylimidodicarbonate [Bioorganic & Medical Chemistry Letters, 1999, 7, (1625-1636)] (8.75 g, 34 tmol) was treated with 9-BBN [Aldrich] (136) at 0 ° C ml, 0.5 M solution in tetrahydrofuran, 2 equivalents, 68 mmol) and the solution stirred at room temperature for 45 minutes. Potassium phosphate (23 ml, 3 M aqueous solution, 2.0 equivalents, 69 mmol) was added carefully and the reaction flask then covered with aluminum foil. A solution of 6-bromo-2-methylbenzothiazole [J. Chem. Soc., 1936, 1225; DE3528032A1] (7.80 g, 34 mmol, 1 equivalent) in dimethylforniamide (50 ml) was added, followed by 1,1'-bis (diphenylphosphino) -ferrocene palladium (II) dichloride-dichloromethane, 1: 1 complex (2.77 g, 3.4 mmol, 0.1 equivalent) and the reaction mixture stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo and the residue purified by column chromatography [SiO 2, pentane: ethyl acetate = 5: 1, then 3: 1] to give the desired product as a clear oil (11.2 g, 84%) delivery-25.

Analysis of ^ -NMR showed that the material had a slight contamination of traces of solvent and 9-BBN.

1 H NMR (CDCl 3, 400 MHz) δκ: 7.81 (1H, d), 7.60 (1H, s), 3.00 7.23 (1H, d), 3.60 (2H, t), 2 77 (3 H, s), 2.71 (2 H, t), 1.97-1.88 (2 H, m), 1.45 (18 H, s); m / z (APCl +): 407 (MH +), 307 (ΜΗ * -Boe).

Preparation 2 102.57 09

70 I

P 1 (tert-butyl) -3- (2-ethyl-1,3-benzothiazol-6-yl) propyl-I

imidodicarbonate I

5 ^ I

° Y ° I

"o. I

10 I

This 2-ethylthiazole intermediate was prepared on an I

manner analogous to that of preparation 1 with 6-bromo-I

2-ethylbenzothiazole [Bull. Soc. Chim. Fr., 1967, 2812-23] I

as the aryl bromide component. I

^ -NMR (dg-DMSO, 400 MHz) 6h: 7.84 (1H, d), 7.64 (1H, s), I

7.26 (1H, d), 3.60 (2H, m), 3.12 (2H, k), 2.73 (2H, t), I

1.07-1.86 (2 H, m), 1.47 (9 H, s), 1.45 (3 H, t); m / z (APCI +): I

20 421 (MH +), 3.21 (MH + -Boc). I

Preparation 3 I

3 - (2-methyl-1,3-benzothiazol-6-yl) propylamine dihydro-1

chloride salt I

25 I

H2 N ^ xxy ~ I

30.2 HCl

N, N-Di-tert-butoxycarbonyl-3- (2-methyl-1,3-benzo)

Thiazol-6-yl) propylamine (11.2 g, 27.5 mmol) was added

102,5709 I

71 dissolved in 1,4-dioxane (30 ml), treated with hydrogen chloride (25 ml, 4M solution in 1,4-dioxane, 100 mmol) and the solution stirred at room temperature for 18 hours. The reaction mixture was evaporated to a white solid which was triturated with a mixture of pentane. and diethyl ether (3: 1) to provide the desired amine bis HCl salt as a white solid (6.05 g, 78%). 1 H NMR (de-DMSO, 400 MHz) 6 h: 7.98 (3 H, brs), 7.85 (1 H, s), 7.81 (1 H, d), 7.31 (1 H, d), 2 78-2.73 (4H, m), 2.75 (3H, 10s),. 1.94-1.85 (2 H, m); m / z (APCI +): 207.

An alternative procedure for the production of the title compound is given in Preparations 4 and 5 below.

15

Preparation 4 3- (2-Methyl-1,3-benzothiazol-6-yl) propylamine dihydrochloride salt (Alternative procedure) 20. Step (1): 3- (2-Methylbenzothiazol-6-yl) acrylonitrile Λ year> -

To a stirred solution of 6-iodo-2-methyl-benzothiazole [WO 2002090443A1] (13.1 g, 47.64 mmol) in 30. DMF (500 ml) was added acrylonitrile (6.27 ml, 95) under Na at room temperature , 28 mmol), then NaOAc (2.86 g, 47.04 mmol), then Pd (0Ac) 2 (1.07 g, 4.76 mmol) and then P (o-tolyl) 3 (2.90 g (9.53 mmol) added. The resulting mixture was 1025709 under Na for ~ 24 hours at 130 ° C and then for 2 days at room temperature

72 I

stirring. The reaction was quenched with water (750 ml) and I

the mixture was then extracted (Et 2 O, 3 x 200 ml). The I

combined organic layers were dried (MgSO 4), gel

filtered and concentrated in vacuo. The resultant I

5 du was purified by flash column chromatography, where I

was eluted with 10: 1 * pentane: EtOAc and then 3: 1 l

pentane -EtOAc, which is the title compound · as an oil I

(9.00 g, 94%) and as a ~ 3: 1 mixture of "trans": "cis" - I

isomers.

10-NMR (6h) 2.85 (3H, m), 5.45 and 5.90 (1H, m), 7.50 (2H, 1

m), 7.90 (2 H, m); I

APCI-MS: 201 (100%) [MH +]. I

Step (2) I

[3- (2-Methylbenzothiazol-6-yl) propyl] carbamic acid tert-I

butyl ester I

J1 ^ - I

20 I

To a stirred solution of 3- (2- I

methyl benzothiazol-6-yl) acrylonitrile (9.00 g, 45.00 mmol) I

in MeOH (1 L), Boc 2 O was added under N 2 at room temperature

25 (19.64 ml, 90.00 mmol) and then NiCl 2 (5.84 g, 45.00 L)

mmol). Portions were added to the resulting mixture

carefully point in ~ 20 minutes NaBIU (13.62 g, 300.00 mmol) I

added. The resulting mixture was added to room I under N 2

temperature stirred for ~ 1.5 hours, filtered through I

30 Arbocel® and concentrated in vacuo. The residue I

was purified by flash column chromatography, wherein I

was eluted with 3: 1 = pentane: EtOAc and then 1: 1 l

pentane: EtOAc, giving the title compound as an oil I

(5.00 g, 36%). I

1025709 I

73-NMR (6 h) 1.40 (9 H, s), 1.85 (1 H, m), 2.65 (1 H, m), 2.75 (1 H, m), 2.90 (3 H, m) ), 3.05 (1H, τη), 3.15 (1H, m), 3.50. (2 H, m), 7.30 (1 H, m), 7.65 (1 H, m), 7.90 (1 H, τη); APCI-MS: 307 (75%) [MH +], 251 (50%), 203 (100%).

5

Step 3) . 3 - (2-Methylbenzothiazole - 6-yl) propylamine dihydrochloro-salt 10 I T v-

H2Ns ^ sAAS

.2HCl

15

To a stirred solution of 3 - (2-methylbenzothiazol - 6-yl] propyl] carbamic acid tert-butyl ester (248 mg, 0.78 mmol) in DOM (10 ml) was added 4N HCl in dioxane (Na) at room temperature ml) and the resulting mixture stirred under Na at room temperature for 4 hours, the solvents were removed in vacuo and the residue triturated with Et 2 O (3 x 10 ml).

The filtrate was concentrated in vacuo to give the title compound as a colorless foam (195 mg, 99%), which, according to 1 H NMR, was of sufficient purity to proceed to the next step without further purification to be used.

Preparation 5.

3- (2-Methyl-1,3-benzothiazol-6-yl) propylamine dihydrochloride salt (Alternative procedure)

Step 1) .

2-13- (2-Methylbenzothiazol-5-yl) propyl) isoindole-1,3-dione 102570Ö

74

I _ y r ^ rN I

5 ° I

I I

I To a slurry of 2- (2-propenyl) -ΙΗ-isoindole-I

1.3 (2H) -dione (18.7 g, 0.1 mol) (allylphthalimide: Journal I

I of Organic Chemistry, (1952), 17, 68-76) in THF (38 ml, 2 l

10 ml / g), a solution (0.5 M) of 9-BBN in THF (240 ml, I

1.2 eq.) In 45 minutes, the temperature being I

I was kept between 0 and 5 ° C. The solution was left

I according to warming up to ambient temperature in one hour and I

I was stirred for an additional 1 hour. A solution of K2 CO3 I

15 (27.6 g, 0.20 mol, 2 eq.) In 50 ml of water was added in 15 minutes

Added, followed by 2-methyl-6-bromo-I

Benzothiazool (20.5 g, 0.09 mol, 0.9 eq.) That was dissolved I

I in DMF (120 ml), and dichloro [1,1 'bis (diphenylphosphino) -1

Ferrocene] palladium (II) dichloromethane adduct (2.4 g, 0.03 l

20 eq.). The reaction mixture was heated for 1 hour

I to 50 ° C and then immediately cooled to ambient temperature

I capture temperature. Water (260 ml, 19 ml / g) and t-BME (560 l

1 ml, 30 ml / g) were added and the solution was added

I stirred. The reaction mixture was filtered through a filter

I aid to remove the solid particles. The phases I

I were separated and the upper organic phase was added

I love. The organic phase was evacuated by vacuum

I filtration concentrated to about 3 ml / g with respect to 1

To the theoretical product. The dark slurry I

I was triturated for 30 minutes at 0 to -5 ° C, driven

I filtered under vacuum and then washed with cooled I

1t-BME (0.4 l, 4.5 ml / g). The resulting solid was obtained

I dried under vacuum at 55 ° C overnight, which I

I 2- [3- (2-methylbenzothiazol-5-yl) propyl) isoindole-1,3-dione I

I (35.5 g, 0.061 mol, 68%). I

I 1025709 I

75. M.p. 126 ° C. . .

-NMR (cU-DMSO, 300 MHz), 5: 1.95-1.98 (m, 2H), 2.72-2.77 (τη, 5H). 3.63 (t, 2H), 7.32 (d, 1H), 7.75 (d, 1H), 7.74-7.83 (m, 5H).

5

Step (2) 3- (2-Methyl-1,3-benzothiazol-6-yl) propylamine dihydrochloride salt:

.2HCl

To a slurry of 2- [3-methyl-benzothiazol-5-yl] propyl) isoindole-1,3-dione (20 g, 0.06 mol) in water (130 ml, 6.5 ml / g) and ethyl alcohol ( 200 ml, 10 ml / g), aqueous methylamine (40 wt / wt solution, 117 g, 1.54 mol, 26 eq.) Was added over 10 minutes, which formed a solution. The solution was stirred at 25 ° C for 18 hours. Dichloromethane (300 ml, 15 ml / g) was added and the reaction mixture stirred for 15 minutes. The phases were separated and. the organic phase was retained. The remaining product was extracted from the aqueous phase with dichloromethane. (100 ml, 5 ml / g). The combined organic phases were washed with 1 M K 2 CO 3 (300 ml, 15 ml / g). The phases were separated and the organic phase was washed with water (300 ml, 15 ml / g). The organic phase was evaporated in vacuo to a small volume and isopropyl alcohol was added, resulting in a final reaction volume of 10 ml / g with respect to 2- [3- (2-methylbenzothiazol-5-yl) propyl] isoindole-1 3-dione. gave. The isopropyl alcohol solution was heated to 70 ° C and concentrated HCl. (12.5 ml, 0.125 mol, 2.1 eq.) 35 was added in 10 minutes. IPA (90 ml, 4.5 ml / g) was evaporated

76

Removed by distillation under atmospheric pressure. The I

I slurry formed by cooling to 25 ° C of the reaction

The solution was filtered off at 0 DEG-5 DEG C. for 1 hour

I was vacuum-washed and washed with cooled IPA (40 ml, 2 l

5 ml / g). The product was under vacuum I overnight

I dried at 55 ° C to give 3- (2-methyl-1,3-benzothiazole)

I-yDpropylamine dihydrochloride salt [12.0 g, 0.058 mol I

I (72%)]; m.p. »215 ° C. I

I 10 Preparation 6 I

3- (2-Ethyl-1,3-benzothiazol-6-yl) -propylamine dihydro-I

Chloride salt I

I rsi # rvrN I

I 15 I

1.2 HCl

I 20 I

This 2-ethylthiazole intermediate was prepared on an I

I manner analogous to that of preparation 3, except that I

I dichloromethane as the first solvent instead of I

I dioxane was used. I

25 ^ -NMR (CDCl 3, 400 MHz) ba: 8.04 (1H, s), 7.95 (1H, d), I

I 7.62 (1H, d), 3.36 (2H, k), 3.02-2.88 (4H, m), 2.10-2.02 I

I C 2 H, m), 1.53 (3 H, t); m / z: 221 (MH +). I

I Preparation 7 I

Tert-Butyl- (2R) -2-methyl-3- [1 - ({[3- (2-methyl-1,3,3-benzo)]

Thiazol-6-yl) propyl] amino] carbonyl) cyclopentyl] propanoate I

35

I 1025709 I

77! H - "o o

A solution of 1- [(2R) -3-tert-butoxy-2-methyl-3-5 oxopropyl] cyclopentanecarboxylic acid [WO 0279143A1] (.6.8 g, 26.5 mmol) in isopropyl acetate (30 ml) was added to. a solution of 1,1'-carbonyldiimidazole (4.76 g, 29.3 mmol, 1.1 equivalent) in isopropyl acetate (60 ml), heating the mixture to 60 ° C for 3 hours and then for 10 minutes. stirred overnight at room temperature. A small volume was removed, evaporated to dryness, and then examined by 1-NMR spectroscopy. This indicated that the reaction proceeded to a conversion of about 90% (Me-doublet for starting acid at δ = 1.15, Me-doublet for acylimidazole at δ> 1.05). Another portion of 1,1'-carbonyldiimidazole (645 mg, 4 mmol, 0.15 equivalent) was added and the mixture stirred for an additional 1 hour at 60 ° C. The reaction mixture was allowed to cool to 40 ° C, triethylamine (4.3 ml, 31 mmol, 1.1 equivalent) and 3 - (2-20 methyl-1,3-benzothiazol-6-yl) propylamine dihydrochloride salt from preparation 3, 4 or 5 (7.1 g, 25.4 mmol, 0.96 equivalent) were added and the mixture was heated to 60 ° C overnight. ; Thin layer chromatography showed that the reaction was not complete. Therefore, another portion of amino dihydrochloride was added (650 mg, 2.3 mmol, 0.09 equivalent), along with triethylamine (330 μΐ, 2.3 mmol, 0.09 equivalent) and the mixture heated to 60 ° overnight c. The reaction mixture was allowed to cool to room temperature and then diluted with water (120 ml and 2 M hydrochloric acid (120 ml). The mixture was extracted with diethyl ether (2 x 400 ml) and the combined extracts were washed with 2 M NaOH (2 x 100 ml), dried (MgSO 4) and evaporated.The resulting oily residue was purified by flash chromatography [SiO 2, 1025709

78 I

methanol in dichloromethane, 0.5% to 1.5%], which is the I

title compound as a clear oil (11.5 g, 98%)

verde. I

-NMR (CDCl 3, 500 MHz) 6h: 7.85 (1H, d), 7.63 (1H, d), 1

5 7.27 (1H, dd), 5.77 (1H, brs), 3.35-3.25 (2H, m) ·, 2.82 I

(3 H, s), 2.77 (2 H, m), 2.33 (1 H, m), 2.03-2.03 (2 H, m), I

1.89 (3 H, m), 1.68-1.55 (5 H, m), 1.45-1.44 (2 H, s), 1.42 I

(9 H, s), 1.10. (3 H, d). I

13 C NMR (CDCl 3, 125 MHz) 5 C: 176.6, 176.6, 166.5, 151.5, I

10 135.8, 126.9, 122.1, 120.8, 80.2, 54.6, 42.4, 39.3, 38.4, I

37.3, 35.0, 33.4, 31.6, 28.0, 24.3, 23.8, 20.0, 19.6. I

.m / z (ES *): 467 MNa +. I

An alternative procedure for the title compound I

is given in the preparation below 8. I

I

Preparation 8 I

tert-Butyl- (2R) -2-methyl-3- [1 - (([3- (2-methyl-1,3-benzo]

thiazol-6-yl) propyl] amino) carbonyl) cyclopentylpropanoate I

(Alternative procedure) I

20 I

Λ I h _ I

γΟγ ^ Ν ^^ ΛΛ ^ I

1 ° O I

25 I

To a slurry of 1,1'-carbonyldiimidazole (154.6 l

g, 1.02 mol) in isopropyl acetate (2.5 ml / g) which is at 60 ° C

was held, a solution of 1 - [(2R) -3-tert-I

Butoxy-2-methyl-3-oxopropyl] cyclopentanecarboxylic acid [WO I

0279143 A1] (283.4 g, 0.93 mol) in isopropyl acetate (2.2 l

ml / g) added for 2 hours under an atmosphere of I

nitrogen gas. The remaining solution of 1 - [(2R) -3-tert-I

butoxy-2-methyl-3-oxopropyl] cyclopentanecarboxylic acid was I

Washed with isopropyl acetate (0.3 ml / g). The solution I

1025709 I

79 was maintained at 60 ° C for an additional 5 hours, stirred and then allowed to cool to room temperature. 3- (2-Methyl-1,3-benzothiazol-6-yl) propylamine dihydrochloride salt from preparation 3, 4 or .5 (259.8 g, 0.93 mol) was added to the solution. Triethylamine (188.3 g, 1.86 mol) was added in 30 minutes. The mixture was brought to reflux in 10 minutes and then kept under reflux for 4 hours, after which it was then allowed to cool to room temperature. The mixture was filtered to remove the insoluble material and washed with isopropyl acetate (2.0 ml / g). The washing liquid was combined with the filtrate. Water (2.0 ml / g) was added and the mixture acidified to pH = 5.0 by adding 5 M hydrochloric acid (375 ml). The mixture was filtered to remove insoluble material. The phases were separated and the upper organic phase was retained. This was then washed with 0.5 M aqueous potassium carbonate solution (238 ml), the phases were separated again and the upper organic layer was retained. This was washed with saturated saline until the pH was less than 9.0. This solution was diluted with isopropyl acetate at 5 ml / g, which was then distilled, after which the solvent was replaced with toluene until a constant volume was maintained at atmospheric pressure. For analytical purposes, a sample could be evaporated to dryness to give the product as an oil (3.89 g, 0.87 mol, 94% yield).

Preparation 9 tert-Butyl-3- [1 - (([3- (2-ethyl-1,3-benzothiazol-6-yl) propyl] amino) carbonyl) cyclopentyl] propanoate 35 1025709

80 I

I η I

* 0 0. I

5 I

1- (3-tert-Butoxy-3-oxopropyl) cyclopentanecarboxylic acid I

[WO 0279143 A1) (180 mg, 0.7 tnmol) was mixed together with I

the 3- (2-ethyl-1,3-benzothiazol-6-yl) propylamine dihydro-1

chloride salt from preparation 6 (180 mg, 0.65 mmol), 1- (3- I

Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (135 L

mg, 0.7 mmol), 1-hydroxybenzotriazole (95 mg, 0.7 mmol) and I

triethylamine (400 μΐ, 2.8 mmol) in dimethylformamide (7 l

ml). The reaction mixture was stirred overnight

at 60 ° C. After cooling to room temperature, the mixture was mixed

15 evaporated in vacuo and the residue diluted with water

(50 ml). The mixture was extracted with diethyl ether I

(3 x 50 ml), the combined organic fractions were I

dried with MgSO 4 and then evaporated. Purification by I

flash chromatography [SiO 2; ethyl acetate in pentane (15 to I

20 gave the desired compound as a light brown I

oil on. I

1 H NMR (CDCl 3, 400 MHz) 6h: 7.87 (1H, d), 7.65 (1H, d), 1

7.26 (1H, dd), 5.63 (1H, brs), 3.30 (2H, k), 3.12 (2H, k), I

2.75 (2H, t), 2.20-2.14 (2H, m), 1.98-1.80 (6H, m), 1.68-1

25 1.58 (5 H, m), 1.45 (3 H, t), 1.42 (9 H, s); I

m / z (ES *): 467 (MNa *), 445 (MH +). I

Alternatively, the title was made to I

using a method analogous to that which I

was described in preparation 7. I

30 I

Preparation 10 I

tert-Butyl- (2R) -2-methyl-3- [1 - (([3- (2-ethyl-1,3-benzothia)

zol-6-yl) propyl] amino) carbonyl) cyclopentyl] propanoate I

35 I

1025709 I

81 i C3 h m ^ - \ v? Y ^^ Ns / s / ^ s x I o o 5

The title compound was prepared by a method analogous to that described for Preparation 9, by coupling the 3- (2-ethyl-1, 3-10 benzothiazol-6-yl) propylamine dihydrochloride salt from Preparation 6 and 1- [(2R) -3-tert-butoxy-2-methyl-3-oxopropyl] -cyclopentanecarboxylic acid [WO0279143A1].

^ -NMR (CDCl 3, 400 MHz) δκ: 7.87 (1 H, d), 7.66 (1 H, s), 7.27 (1 H, m), 5.77 (1 H, brm), 3.29 (2 H, m), 3.13 (2 H, k), 2.77 (2 H, m), 2.31 (1 H, m), 2.08-1.96 (2 H, m), 1, 70-1.43 (10 H, m), 1.45 (3 H, t), 1.43 (9 H, s), 1.10 (3 H, d); m / z (ES +): 481 (MNa +), 459 (MH +).

Preparation 11 20 1- (2-tert-Butoxycarbonylethyl) cyclohexanecarboxylic acid "Ύ -

1 O O

Cyclohexanecarboxylic acid (2.89 g, 22.6 mmol) was dissolved in THF (30 mL) and this solution was added at such a rate to a solution of lithium diisopropylamide cooled to -15 ° C (2M in THF / n-heptane / ethylbenzene, Aldrich) (24.3 ml, 48.6 mmol), that the temperature. remained below 0 ° C. The reaction mixture was then stirred at 0 ° C for 2.5 hours before being cooled again to -15 ° C and tert-butyl 3-bromopropionate (525259)

82

I g, 23.9 tmmol) in THF (50 ml) at such a rate was I

I added that the temperature remained below 0 ° C. I

I the reaction mixture came to room temperature and I

I was stirred overnight before the reaction was complete

I was broken by the addition of 2M HCl and the mixture became I

I extracted with 2 x 200 ml of ethyl acetate. The combined I

I the organic layers were dried (MgSO 4) and then I

I evaporated to an orange oil. Purification by flash I

Chromatography iSiO 2, dichloromethane / methanol / NH 3 (s.d. I

I 10 = 0.880) (97: 3: 0.5)) gave the title compound as an I

I gummy oil (800 mg, 14%). I

1-NMR (CDCl 3, 400 MHz) δκ: 2.22 (2H, m), 2.07-2.00 (2H, 1

M), 1.86 (2 H, m), 1.45-1.16 (8 H, m), 1.41 (9 H, s). I

I m / z (ES +): 255 (M-H4) I

I 15 I

I Preparation 12 I

Tert-Butyl-3- (1 - {[3- (2-ethyl-1,3-benzothiazol-6-yl) pro-]

Pyl) carbamoyl} cyclohexyl propanoate I

I 20 I

I v C J H I

I ^ ° s ^ sX, N I

I> 00 I

I 25 I

The title compound was prepared using an I

I method analogous to that described I

I for preparation 9 with 3- (2-ethyl-1,3-benzothiazole-6-I

Dopopylamine dihydrochloride salt from preparation 6 together I

30 with .1 - [(2R) -3-tert-butoxy-2-methyl-3-l

Oxopropyl) cyclohexanecarboxylic acid from preparation 11. I

^ -NMR (CDCl3, 400 MHz) δκ: 7.88 (1H, d), 7.65 (1H, s), I

7.27 (1H, m), 5.65 (1H, brm), 3.34 (2H, m), 3.14 (2H, k), I

2.77 (2H, m), 2.15 (1H, m), 1.95-1.27 (14H, m), 1.45 (3H, I)

35t), 1.41 (9H, s); I

1025709 I

83 m / z (ES +): 481 (MNa +), 459 (MH +).

Preparation 13 tert-Butyl 3- (1- ([3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl) cyclohexyl) propanoate counting compound was prepared by a method analogous to that described for preparation 9, with 3- (2-methyl-1,3-benzothiazol-e-yl) propylamine dihydrochloride salt from preparation 3, 4 or 5 together with 1- [(2R) -3-tert-butoxy-2-methyl-3-oxopropyl] cyclohexane carboxylic acid from preparation 11.

1 H NMR (CDCl 3, 400 MHz) δκ: 7.85 (1H, d), 7.64 (1H, d), 7.27 (hidden) (1H, dd), 5.65 (1H, brm), 3.33 (2 H, m), 2.81 (3 H, s), 2.77 (2 H, m), 2.18-2.13. (2H, m), 1.93-1.79 (4H, m), 1.74-1.68 (2H, m), 1.60-1.23 (7H, m), 1.40 (9H , 25 s); m / z (ES +): 467 (MNa +), 445 (MH4).

Preparation 14 tert-Butyl 3- (1- ([3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl) cyclopentyl) propanoate 35, 1025709

84 I

The title compound was prepared using an I

method analogous to that described I

for preparation 9 by coupling 3- (2-methyl-1,3,3)

Benzothiazol-6-yl) propylamine dihydrochloride salt from b

line 3, 4 or 5 and tert-butyl 3- (1-carboxycyclopentyl) -1

propanoate [WO 0279143A1]. I

1 H NMR (CDCl 3, 400 MHz) δκ: 7.85 (1H, d), 7.62 (1H, d), 1

7.25 (1 H, m), 5.63 (1 H, brm), 3.29 (2 H, m), 2.81 (3 H, s), I

10 2.75 (2H, m), 2.19-2.15 (2H, m), 2.00-1.36 (11H, m), 1.42 l

(9 H, s); I

m / z (APCI +): 431 (MH +). I

1025709 I

Claims (16)

A compound of formula (I) 5 · r-Wn? i) H 'or W 10. O (I) wherein; R1 is H or CH3, R 2 is C 1 -C 6 alkyl and η is 1 or 2, a tautomer thereof or a pharmaceutically acceptable salt, solvate or polymorph of the compound or tautomer. 20 2.. A compound according to claim 1, wherein η is 1. A compound according to claim 1 or claim 2, wherein R 1 is hydrogen. A compound according to claim 1 or claim 2, wherein R 1 is methyl. A compound according to any one of claims 1-4, wherein R 2 is methyl. A compound according to any one of claims 1-4, wherein R 2 is ethyl. 7. A compound according to claim 1, which is selected from: (R) -2-Methyl-3- (1- {[3- (2-methyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl propanoic acid (Example 1). 3- (1 - {[3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] -35-carbamoyl-cyclopenty-propanoic acid (Example 2), 1025709 86 I (R) -2-Methyl-3- (1 - {[3- (2-ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclopentyl) propanoic acid (Example 4) and 3- (1- {[3- (2-Ethyl-1,3-benzothiazol-6-yl) propyl] carbamoyl} cyclohexyl) propanoic acid (Example 3). I A pharmaceutical composition comprising a compound of formula (I) as claimed in any one of claims 1-7, or pharmaceutically acceptable salts, solvates or polymorphs I thereof, and a pharmaceutically acceptable (a) r (e) - comprises thinner or carrier. A compound of formula (I) according to any of claims 1-7, or a pharmaceutically acceptable salt, solvate or polymorph thereof, for use as a medicament. A method for the treatment of a disorder or condition in which it is known, or can be demonstrated, that inhibition of NEP produces a beneficial effect in a mammal, which comprises administering to a mammal a mammal therapeutically effective amount of a compound of formula (I) according to any one of claims 1-7, or a pharmaceutically acceptable salt, solvate or polymorph thereof. I 11. Use of a compound of formula (I) according to one of claims 1-7, or a pharmaceutically acceptable salt, solvate or polymorph thereof, in the preparation of a medicament for the treatment I of a disorder or disorder in which it is known or can be demonstrated that inhibition of NEP produces a beneficial effect. I 12. A compound according to claim 9, method according to H claim 10 or use according to claim 11, wherein I the disorder or condition is selected from hypertenesis, essential hypertension, pulmonary hypertension, I secondary hypertension, isolated systolic hypertension, hypertension associated with diabetes, hypertension associated with atherosclerosis and renovascular hypertension, peripheral vascular disease, cardiac insufficiency, angina, renal insufficiency, acute renal insufficiency, cyclic edema, disease of Meniere, hyperaldosteroneemism (primary) hypercalciuria, stroke, glaucoma, obesity, metabolic diseases, metabolic syndrome, diabetes, impaired glucose tolerance, diabetic retinopathy, diabetic neuropathy, menstrual disorders, premature labor pains, preeclampsia, endometryriosis and reproductive disorders, male and female infertility, polycystic ovarian syndrome, m failure implantation; asthma, inflammation, leukemia, pain, pain, depression, drug use, cirrhosis, epilepsy, affective disorders, dementia and geriatric confusion, gastrointestinal disorders, diarrhea, irritable bowel syndrome, wound healing, diabetic and venous ulcers and bedsores, septic shock, heartburn, hyperreninism, cystic fibrosis, restenosis, atherosclerosis, female sexual dysfunction (FSD), sexual arousal disorder, female sexual arousal disorder (FSAD), male sexual dysfunction (male sexual dysfunction, MSD), male erectile dysfunction (MED), hypoactive sexual desire disorder, orgasmic disorder and sexual pain disorder. A compound, method or use according to claim 12, wherein the disorder or condition is selected from female sexual dysfunction (FSD), sexual arousal disorder, female sexual arousal steamis (FSAD), male sexual dysfunction (MSD), male erectile dysfunction ( MED), hypoac-1025709 88 Iive sexual pleasure disorder, orgasmic disorder and I sexual pain disorder. I A compound, method or use according to claim 13, wherein the disorder or condition is selected from female sexual dysfunction (PSD), female sexual arousal disorder (FSAD), male sexual dysfunction (MSD) and male erectile I dysfunction (MED). I 1025709 I